Comparing X-Ray Film and Digital Imaging Systems

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X-ray film has long been considered the gold standard of sensitivity in western blotting, but the broad dynamic range enabled by digital imaging systems allows for accurate quantitation of a range of signal intensities. With recent technological advances, can researchers finally have the best of both worlds?

Advantages of Digital Imaging Systems

Key benefits to using this technology include:

  • Wide dynamic range = > 4 logs
  • Quantitative
  • Digital documentation
  • Environmentally friendly — no chemicals!
  • No costly consumables (film)
  • Initial capital investment typically recovered
    in as little as 1 year

See the Data

The sensitivity of film, with a broad dynamic range and superior signal-to-noise.

Comparison of sensitivity between the ChemiDoc MP Imaging System and film. A. Western blot analysis of LacI expression was conducted using 2x serial dilutions (starting at 0.31 μg protein) of E. coli cell lysate. The membranes were either imaged on the ChemiDoc MP Imaging System for 30 sec or exposed to film for 30 sec. B. Band intensities, normalized to lane 7 background, illustrate the ability of the ChemiDoc MP Imaging System to detect low signal bands at the same exposure time as film. The red boxes represent the limited linear dynamic range of film. ChemiDoc MP Imaging System, 30 sec (   ); film, 30 sec (   ).

Side-by-side comparison between the ChemiDoc Imaging System and film. Levels of the three isoforms of the pro-apoptotic protein Bim were measured in various cell lines using western blot analysis. The membranes were either imaged on the ChemiDoc Imaging System for 30 sec or exposed to film for 30 sec to compare detection sensitivities. As shown by the overlay graph, the ChemiDoc Imaging System was better able to detect faint protein bands than film.

 

ChemiDoc MP Imaging System: Imagine No Compromises

With the ChemiDoc MP Imaging System, the superior quantitation of digital imaging meets the sensitivity of film. This best-in-class system images and analyzes gels and western blots. It is designed for multiplex fluorescent western blotting, chemiluminescence detection, general gel documentation, and stain-free technology imaging needs.

Learn More

ChemiDoc Imaging System

Better Options. Better Results.

Stain-Free
Total Protein Normalization

Learn how to properly design a quantitative western blotting experiment and faithfully compare protein expression levels, without concern for changing or overexpressed HKP’s.
See Data »

Chemiluminescence
Replaces Film

Save money on consumables and reduce environmental waste without compromising on performance. Get the sensitivity of film and the convenience of digital documentation.
See Data »

Multiplex Fluorescence
3 Colors

Save sample, avoid the errors commonly associated with stripping, reprobing, and cutting blots and answer more complex biological questions by optimizing your experiments to fluorescence.
See Data »

Publication Requirements

Need another reason to get out of the darkroom?

Several journals have recently revised their guidelines for publication, providing guidance around reproducibility and quantitation.

Defining the New Normal in Quantitative Western Blot Data

Learn why total protein normalization (TPN) is a superior method for quantitative western blotting analysis.

Publication requirements are changing. Are your blots ready?

  • 1

    "House-keeping proteins should not be used for normalization without evidence that experimental manipulations do not affect their expression"*

    Solution: Use Total Protein Normalization (TPN) instead of house-keeping proteins

    • Bio-Rad's stain-free workflow makes TPN easy! This technology is proven with over 100 publications, allows for improved accuracy and will make your data more reliable
  • 2

    "Methods including detection of enhanced chemiluminescence using X-ray film have a very limited dynamic range"*

    Solution: Use an imaging system with up to 4 logs of dynamic range

    • Bio-Rad's ChemiDoc System — an imager with the sensitivity of film
    • Simple image generation and quantitation using Image Lab Software
  • 3

    "A description of the data supporting the specificity of all antibodies is required"*

    Solution: Fully validated antibodies

    • Bio-Rad's PrecisionAb Antibodies offer: documented specificity and validation protocols

* Revised guidelines for authors from The Journal of Biological Chemistry

 

Total Protein Normalization provides accurate quantitation of target proteins.

Normalizing blots with housekeeping proteins (HKPs) requires time, money, and additional sample to validate HKP expression and linearity. Total protein normalization using stain-free technology and the ChemiDoc or ChemiDoc MP System allows normalization across a wide dynamic range. Get better, more publishable data without lengthy optimization.

Linearity comparison of stain-free total protein measurement and immunodetection of three housekeeping proteins in 10–50 µg of HeLa cell lysate.

Learn more about Stain-free Technology

Bio-Rad's V3 Western Workflow Incorporates TPN for Normalization of Western Blot Data

V3 Western Workflow includes:

  • Stain-free visualization of total proteins on gel after electrophoresis
  • Quantification of total proteins in a lane after transfer
  • Normalization of bands to total proteins by specialized software

Find out why these 3 steps are crucial to generating quantitative western blot data.

Learn More  Request a Demo

FEATURED PRODUCTPrecisionAb Validated Primary Antibodies
for Western Blotting

  • Comprehensive selection of antibodies for cancer and neuroscience research
  • Highly specific and sensitive
  • Validated against up to 12 lysates
  • Stringent QC criteria for superior consistency

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Design your Western Blotting Experiments with Confidence

As an editor of several journals, Professor Aldrin V. Gomes routinely observes western blots that are not properly conducted. The generally low quality of western blotting data has led scientists to question whether we can trust protein quantification when using western blots. Reducing errors and misinterpretations requires standardized procedure and proper controls. Dr. Gomes discusses the following key factors for high-confidence western blot data:

  1. Quality of the primary and secondary antibodies and the blocking reagent
  2. Proper target protein controls
  3. Choice of total protein loading control vs. housekeeping protein loading control to normalize the target protein measurement

Prof. Gomes also presents tips to reduce the overall time required for first-rate western blots, and how to minimize the pain of going through a long procedure.

 
  • Reliable Quantification of Western Blot Data

    Based on the article "A Defined Methodology for Reliable Quantification of Western Blot Data". Dr. S.C Taylor provides a direct comparison between results generated from camera-based imaging technologies and film using identical blots. Learn common pitfalls associated with film use and densitometric analysis.

    This video is based on the article published by S.C. Taylor et al. entitled "A Defined Methodology for Reliable Quantification of Western Blot Data" (DOI 10.1007/s12033-013-9672-6).

  • Designing a Quantitative Western Blot Experiment to Avoid Common Pitfalls

    The quantitative western blot methodology is described with a focus on solid experimental design and data analysis to achieve reproducible, publication quality results in the shortest time frame.

    This video is based on the article published by Drs. Taylor and Posch entitled: "The Design of a Quantitative Western Blot Experiment".

  • Quantitative Western Blotting: Optimization is the Key

    Dr. S.C. Taylor details the common errors associated with performing quantitative western blots and how they can be addressed through a rigorous optimization methodology. Each step of the quantitative western blot workflow is discussed with a focus on optimization to assure quantitative, reproducible final data that reflect the experimental conditions.

Testimonial

Leukemia researcher Ernesto Diaz-Flores (University of California, San Francisco) describes how the ChemiDoc Imaging System helped him improve his western blot imaging, enabling him to identify relevant therapeutic targets. An avid photographer, Prof. Diaz-Flores also talks about the links between his art and his science and technologies that help him in both fields.

Read Full Article

Resources

FEATURED WEBINARUnderstand the Factors That are Crucial to Successful Western Blotting

Successful Western Blotting

Join Bio-Rad western blotting expert Kenneth Oh and his guest speakers for a webinar series that explores the many factors that go into the design and execution of successful and repeatable western blots. Chemiluminescence or fluorescence, qualitative or quantitative blot, not only will we discuss the how-to's, we'll take a deeper dive and discuss the why's.

View Webinar


  • Image Lab Touch Software Tutorials

    Image Lab Touch Software Resources

    Bio-Rad provides instructional videos for Image Lab Touch Software 2.4, covering the primary applications of the ChemiDoc MP Imaging System. Topics include, logging in, Smart Tray Technology, capturing your image, and exporting data.

  • Image Lab Software Tutorials

    Image Lab Software Resources

    Bio-Rad provides tutorials for basic acquisition and advanced analysis use of Image Lab Software 6.0.1. Topics include, densitometric analysis, molecular weight, normalization, purity assessment, and more!

 

Application Guides

  • Western Blot Dr. Troubleshooting Guide

    Western Blot Doctor Troubleshooting Guide

    Our self-help troubleshooting guide covers solutions to many common and not-so-common western blotting issues and helps your blots look their best.

  • Protein Blotting Guide

    Protein Blotting Guide

    (PDF 7.9 MB)
    Details on blotting technology, available products, and methods, plus tips, techniques, and troubleshooting.

  • Electrophoresis Guide

    Electrophoresis Guide

    (PDF 6.4 MB)
    A guide to polyacrylamide gel electrophoresis and protein detection, including theory, product selection, protocols, and more.

 

Documents Library

 

Publications

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    Mini-PROTEAN TGX Stain-Free Precast Gels for polyacrylamide gel electrophoresis (PAGE) allow for fast run times, high transfer efficiency, and rapid detection of proteins with stain-free enabled imagers.

  • PrecisionAb Validated Western Blotting Antibodies

    PrecisionAb Validated Western Blotting Antibodies

    These premium antibodies are lab-validated using strict testing criteria to ensure superior performance in western blotting detection.

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PNK1ECTU86LJ [x-forwarded-proto] = [https] [x-forwarded-port] = [443] [x-forwarded-for] = [176.97.36.93, 10.232.19.88] [accept] = [*/*] [seourl] = [/en-us/applications-technologies/comparing-x-ray-film-digital-imaging-systems] [x-amzn-trace-id] = [Root=1-5d83ba9c-99cc1e6c2f03f2bc410fc878] [x-forwarded-server] = [lsds-prod-s.br.aws-livesite.io] [x-forwarded-host] = [www.bio-rad.com] [x-query-string] = [ID=PNK1ECTU86LJ] [host] = [10.232.1.21:1776] [x-request-uri] = [/en-us/applications-technologies/comparing-x-ray-film-digital-imaging-systems] [connection] = [Keep-Alive] [accept-encoding] = [deflate, gzip] [user-agent] = [Mozilla/5.0 (Windows NT 6.0; rv:34.0) Gecko/20100101 Firefox/34.0] AppTech/AppTechDetails pageStyleKey internet/solutions_sub applications-technologies/comparing-x-ray-film-digital-imaging-systems LSR PNK1ECTU86LJ Comparing X-Ray Film and Digital Imaging Systems Comparing X-Ray Film and Digital Imaging Systems <div class="pagenav"><img src="/webroot/web/images/opt/prodnav-arrow-top.png" alt="" /> <table border="0"> <tbody> <tr> <td class="pagenavhead"> <h3>On This Page</h3> </td> <td class="pagenavfirst"><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Page Nav&amp;tgp_pcp_link=Advantages of Digital Imaging Systems','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="#advantages">Advantages of Digital Imaging Systems</a></td> <td><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Page Nav&amp;tgp_pcp_link=ChemiDoc Imaging System','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="#ChIS">ChemiDoc Imaging System</a></td> <td><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Page Nav&amp;tgp_pcp_link=Publication Requirements','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="#publication">Publication Requirements</a></td> <td><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Page Nav&amp;tgp_pcp_link=Designing Experiments','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="#designing">Designing Experiments</a></td> <td><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Page Nav&amp;tgp_pcp_link=Resources','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="#resources">Resources</a></td> <td class="pagenavlast"><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Page Nav&amp;tgp_pcp_link=Related Products','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="#related">Related Products</a></td> </tr> </tbody> </table> </div> <div class="opt-module opt-module-first opt-module-noborder"> <div class="corner-box corner-box-promo linkpointer-blank"><img src="/webroot/web/images/general/global/english/have-questions-icon.png" border="0" alt="Consult with an Specialist" style="width: 50px; height: auto;" /> <p><strong>Need help with imaging?</strong></p> <a href="https://info.bio-rad.com/ww-image-lab-twas.html" class="bttn-block no-target-style" target="_blank" rel="noopener noreferrer"><span class="bttn-orange">Consult with a Specialist</span></a></div> <p>X-ray film has long been considered the gold standard of sensitivity in western blotting, but the broad dynamic range enabled by digital imaging systems allows for accurate quantitation of a range of signal intensities. With recent technological advances, can researchers finally have the best of both worlds?</p> </div> <div><a name="advantages"></a></div> <div class="opt-module bg-f0f9f8 opt-module-noborder"> <h2 class="nextp">Advantages of Digital Imaging Systems</h2> <iframe src="https://www.youtube.com/embed/pzIRIrhgG2M?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" height="245px" width="437px" _fsrb="true" class="right"></iframe> <p><strong>Key benefits to using this technology include:</strong></p> <ul> <li>Wide dynamic range = &gt; 4 logs</li> <li>Quantitative</li> <li>Digital documentation</li> <li>Environmentally friendly &mdash; no chemicals!</li> <li>No costly consumables (film)</li> <li>Initial capital investment typically recovered <br />in as little as 1 year</li> </ul> <div class="corner-box bg-white top-margin"> <h2 class="no-bttm-pddng">See the Data</h2> <h3 class="opt-subhead30">The sensitivity of film, with a broad dynamic range and superior signal-to-noise.</h3> <center><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/imaging_and_analysis/technology_detail/low-signal-detection-film-data-sensitivity.png" alt="" class="top-margin" /></center> <p class="caption topmargin-1em"><strong>Comparison of sensitivity between the ChemiDoc MP Imaging System and film. A.</strong> Western blot analysis of LacI expression was conducted using 2x serial dilutions (starting at 0.31 &mu;g protein) of E. coli cell lysate. The membranes were either imaged on the ChemiDoc MP Imaging System for 30 sec or exposed to film for 30 sec. <strong>B.</strong> Band intensities, normalized to lane 7 background, illustrate the ability of the ChemiDoc MP Imaging System to detect low signal bands at the same exposure time as film. The red boxes represent the limited linear dynamic range of film. ChemiDoc MP Imaging System, 30 sec ( <span class="bar-graph-key bg-95cb45">&nbsp;</span> ); film, 30 sec ( <span class="bar-graph-key bg-184a74">&nbsp;</span> ).</p> <center><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/imaging_and_analysis/technology_detail/reveal-faint-bands-film-data-sensitivity.png" alt="" class="top-margin" /></center> <p class="caption topmargin-1em"><strong>Side-by-side comparison between the ChemiDoc Imaging System and film.</strong> Levels of the three isoforms of the pro-apoptotic protein Bim were measured in various cell lines using western blot analysis. The membranes were either imaged on the ChemiDoc Imaging System for 30 sec or exposed to film for 30 sec to compare detection sensitivities. As shown by the overlay graph, the ChemiDoc Imaging System was better able to detect faint protein bands than film.</p> </div> &nbsp;<br clear="all" /> <iframe src="https://www.youtube.com/embed/xAUFWRONV_U?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" mce_src="https://www.youtube.com/embed/nVsLxeT8Reg?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" height="245px" width="437px" _fsrb="true" class="left"></iframe> <h3 class="opt-subhead">ChemiDoc MP Imaging System: Imagine No Compromises</h3> <p>With the ChemiDoc MP Imaging System, the superior quantitation of digital imaging meets the sensitivity of film. This best-in-class system images and analyzes gels and western blots. It is designed for multiplex fluorescent western blotting, chemiluminescence detection, general gel documentation, and stain-free technology imaging needs.</p> <p><a class="bttn-block no-target-style" onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Product Spotlight&amp;tgp_pcp_link=Learn More','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/en-us/category/imaging-systems-software?ID=f97d1fa1-16ff-4c73-9eae-ef74be3a3f95"><span class="bttn-orange">Learn More</span></a></p> </div> <div><a name="ChIS"></a></div> <div class="opt-module opt-module-noborder bg-f0f9f8"> <h2>ChemiDoc Imaging Systems</h2> <h3 class="opt-subhead30 top-margin-neg">Complete Capabilities: Superior Images</h3> <div id="interactiveRibbonOuter"> <div class="ribbonThird" style="background-image: url('/webroot/web/images/lsr/global/english/solutions/technologies/Fluorescent-Western-Blotting/stain-free-square.png'); background-repeat: no-repeat;"> <div class="ribbonThirdInner"> <div class="ribbonHeader"> <div class="ribbon2"> <div class="ribbon2Center"> <h4 class="gy05">Stain-Free<br /><small>Total Protein Normalization</small></h4> </div> </div> <div class="ribbon2Right"></div> </div> <div class="ribbonDesc"> <p>Learn how to properly design a quantitative western blotting experiment and faithfully compare protein expression levels, without concern for changing or overexpressed HKP&rsquo;s. <br /><a href="/en-sg/applications-technologies/total-protein-normalization?ID=PODYJQRT8IG9">See Data &raquo;</a></p> </div> </div> </div> <div class="ribbonThird" style="background-image: url('/webroot/web/images/lsr/global/english/solutions/technologies/Fluorescent-Western-Blotting/chemiluminescence.jpg'); background-repeat: no-repeat;"> <div class="ribbonThirdInner"> <div class="ribbonHeader"> <div class="ribbon2"> <div class="ribbon2Center"> <h4 class="gy05">Chemiluminescence<br /><small>Replaces Film</small></h4> </div> </div> <div class="ribbon2Right"></div> </div> <div class="ribbonDesc"> <p>Save money on consumables and reduce environmental waste without compromising on performance. Get the sensitivity of film and the convenience of digital documentation. <br /><a href="/en-sg/applications-technologies/comparing-x-ray-film-digital-imaging-systems?ID=PNK1ECTU86LJ">See Data &raquo;</a></p> </div> </div> </div> <div class="ribbonThird" style="background-image: url('/webroot/web/images/lsr/global/english/solutions/technologies/Fluorescent-Western-Blotting/bio-rad-fluorescence.jpg'); background-repeat: no-repeat;"> <div class="ribbonThirdInner"> <div class="ribbonHeader"> <div class="ribbon2"> <div class="ribbon2Center"> <h4 class="gy05">Multiplex Fluorescence<br /><small>3 Colors</small></h4> </div> </div> <div class="ribbon2Right"></div> </div> <div class="ribbonDesc"> <p>Save sample, avoid the errors commonly associated with stripping, reprobing, and cutting blots and answer more complex biological questions by optimizing your experiments to fluorescence. <br /><a href="/en-sg/applications-technologies/fluorescent-western-blotting?ID=PJW4ANE08O1Y">See Data &raquo;</a></p> </div> </div> </div> </div> </div> <div><a name="publication"></a></div> <div class="opt-module opt-module-noborder"> <h2 class="nextp">Publication Requirements</h2> <h3 class="opt-subhead">Need another reason to get out of the darkroom?</h3> <p>Several journals have recently revised their guidelines for publication, providing guidance around reproducibility and quantitation.</p> <ul class="list-set list-four-col topmargin-1em"> <li><center><a href="https://www.nature.com/authors/policies/image.html" target="_blank" class="no-target-class" rel="noopener noreferrer"><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/imaging_and_analysis/technology_detail/website-nature-4col-th.png" alt="" /></a> <h3><a href="https://www.nature.com/authors/policies/image.html" target="_blank" class="no-target-class" rel="noopener noreferrer">Nature</a></h3> </center></li> <li><center><a href="https://science.sciencemag.org/content/346/6210/679.full" target="_blank" class="no-target-class" rel="noopener noreferrer"><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/imaging_and_analysis/technology_detail/website-science-4col-th.png" alt="" /></a> <h3><a href="https://science.sciencemag.org/content/346/6210/679.full" target="_blank" rel="noopener noreferrer">Science</a></h3> </center></li> <li><center><a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4705984/" target="_blank" class="no-target-class" rel="noopener noreferrer"><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/imaging_and_analysis/technology_detail/website-ncbi-4col-th.png" alt="" /></a> <h3><a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4705984/" target="_blank" rel="noopener noreferrer">National Center for Biotechnology Information</a></h3> </center></li> <li><center><a href="https://www.jbc.org/site/misc/ifora.xhtml" target="_blank" class="no-target-class" rel="noopener noreferrer"><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/imaging_and_analysis/technology_detail/website-jbc-4col-th.png" alt="" /></a> <h3><a href="https://www.jbc.org/site/misc/ifora.xhtml" target="_blank" rel="noopener noreferrer">Journal of Biological Chemistry</a></h3> </center></li> </ul> <div class="corner-box corner-box-tpn linkpointer-blank"> <h3><a href="https://www.bioradiations.com/defining-the-new-normal-in-quantitative-western-blot-data" target="_blank" rel="noopener noreferrer">Defining the New Normal in Quantitative Western Blot Data</a></h3> <p>Learn why total protein normalization (TPN) is a superior method for quantitative western blotting analysis.</p> </div> <h3 class="opt-subhead30 top-margin">Publication requirements are changing. Are your blots ready?</h3> <ul class="flex-container flex-3-col flex-col-stroke leaf-border flex-pub-req"> <li> <h3><span>1</span></h3> <p><strong>"House-keeping proteins should not be used for normalization without evidence that experimental manipulations do not affect their expression"*</strong></p> <p><strong>Solution:</strong> Use Total Protein Normalization (TPN) instead of house-keeping proteins</p> <ul class="no-list-pddng disc-list"> <li>Bio-Rad's stain-free workflow makes TPN easy! This technology is proven with over 100 publications, allows for improved accuracy and will make your data more reliable</li> </ul> </li> <li> <h3><span>2</span></h3> <p><strong>"Methods including detection of enhanced chemiluminescence using X-ray film have a very limited dynamic range"*</strong></p> <p><strong>Solution:</strong> Use an imaging system with up to 4 logs of dynamic range</p> <ul class="no-list-pddng disc-list"> <li>Bio-Rad's ChemiDoc System &mdash; an imager with the sensitivity of film</li> <li>Simple image generation and quantitation using Image Lab Software</li> </ul> </li> <li> <h3><span>3</span></h3> <p><strong>"A description of the data supporting the specificity of all antibodies is required"*</strong></p> <p><strong>Solution:</strong> Fully validated antibodies</p> <ul class="no-list-pddng disc-list"> <li>Bio-Rad's PrecisionAb Antibodies offer: documented specificity and validation protocols</li> </ul> </li> </ul> <p class="caption top-margin">* Revised guidelines for authors from <a href="http://www.jbc.org/site/misc/ifora.xhtml#blots" target="_blank" rel="noopener noreferrer">The Journal of Biological Chemistry</a></p> <div class="dotteddivider dotteddividerdrk dotteddivider-less-top">&nbsp;</div> <h3 class="opt-subhead">Total Protein Normalization provides accurate quantitation of target proteins.</h3> <p>Normalizing blots with housekeeping proteins (HKPs) requires time, money, and additional sample to validate HKP expression and linearity. Total protein normalization using stain-free technology and the ChemiDoc or ChemiDoc MP System allows normalization across a wide dynamic range. Get better, more publishable data without lengthy optimization.</p> <img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/imaging_and_analysis/technology_detail/acurate-tpn.png" class="topmargin-1em" alt="" /> <p class="caption topmargin-1em"><strong>Linearity comparison of stain-free total protein measurement and immunodetection of three housekeeping proteins in 10&ndash;50 &micro;g of HeLa cell lysate.</strong></p> <div class="corner-box bg-white no-bttm-pddng topmargin-1em corner-box-stainfree"> <h3 class="opt-subhead30">Learn more about Stain-free Technology</h3> <ul class="list-set list-two-col list-set-th"> <li><a href="https://www.bioradiations.com/trends-in-protein-separation-and-analysis-the-advance-of-stain-free-technology/" target="_blank" rel="noopener noreferrer"><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/imaging_and_analysis/technology_detail/" alt="" /></a> <h3><a href="https://www.bioradiations.com/trends-in-protein-separation-and-analysis-the-advance-of-stain-free-technology/" target="_blank" rel="noopener noreferrer">Trends in Protein Separation and Analysis &mdash; the Advance of Stain-Free Technology</a></h3> <p>This article focuses on the theory and concepts surrounding stain-free technology as well as other traditional methods of protein staining.</p> </li> <li> <h3><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=AppTech&amp;tgp_pcp_link=Stain-Free Imaging Technology','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/en-us/applications-technologies/stain-free-imaging-technology">Stain-Free Imaging Technology</a></h3> <p>Visualize proteins on gels and blots without staining and destaining steps, and use total protein normalization for quantitative western blots.</p> </li> </ul> </div> <h3 class="opt-subhead30">Bio-Rad's V3 Western Workflow Incorporates TPN for Normalization of Western Blot Data</h3> <iframe src="https://www.youtube.com/embed/RWsDtTqDW_o?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" height="245px" width="437px" class="right"></iframe> <p>V3 Western Workflow includes:</p> <ul> <li>Stain-free visualization of total proteins on gel after electrophoresis</li> <li>Quantification of total proteins in a lane after transfer</li> <li>Normalization of bands to total proteins by specialized software</li> </ul> <p>Find out why these 3 steps are crucial to generating quantitative western blot data.</p> <p><a class="bttn-block linkgeneration" onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=V3&amp;tgp_pcp_link=Learn more','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/en-us/product/v3-western-workflow"><span class="bttn-orange">Learn More</span></a> &nbsp;<a class="bttn-block no-target-style" onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=V3&amp;tgp_pcp_link=Request a Demo','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="https://info.bio-rad.com/ww-V3-Central-Umbrella-btwas.html" target="_blank" rel="noopener noreferrer"><span class="bttn-green">Request a Demo</span></a></p> </div> <div class="opt-module white opt-module-precisionab-feature linkpointer"> <h2 class="nextp white"><span class="featuredtxt">FEATURED PRODUCT</span>PrecisionAb Validated Primary Antibodies<br />for Western Blotting</h2> <ul class="list-orange-arrow"> <li class="white">Comprehensive selection of antibodies for cancer and neuroscience research</li> <li class="white">Highly specific and sensitive</li> <li class="white">Validated against up to 12 lysates</li> <li class="white">Stringent QC criteria for superior consistency</li> </ul> <p><a class="bttn-block" onmousedown="dcsMultiTrack('DCS.dcsqry','? tgp_pcp_link_group=PrecisionAb Validated Western Blotting Antibodies&amp;tgp_pcp_link=Learn more','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999'); return false;" href="/en-us/product/precisionab-validated-western-blotting-antibodies"><span class="bttn-orange">Find Your Antibody</span></a></p> </div> <div><a name="designing"></a></div> <div class="opt-module opt-module-noborder no-bttm-pddng"> <h2>Design your Western Blotting Experiments with Confidence</h2> <iframe src="https://www.youtube.com/embed/zrfAHAo4xT0?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" height="245px" width="437px" _fsrb="true" class="right"></iframe> <p>As an editor of several journals, Professor Aldrin V. Gomes routinely observes western blots that are not properly conducted. The generally low quality of western blotting data has led scientists to question whether we can trust protein quantification when using western blots. Reducing errors and misinterpretations requires standardized procedure and proper controls. Dr. Gomes discusses the following key factors for high-confidence western blot data:</p> <ol> <li>Quality of the primary and secondary antibodies and the blocking reagent</li> <li>Proper target protein controls</li> <li>Choice of total protein loading control vs. housekeeping protein loading control to normalize the target protein measurement</li> </ol> <p>Prof. Gomes also presents tips to reduce the overall time required for first-rate western blots, and how to minimize the pain of going through a long procedure.</p> <div class="dotteddivider dotteddividerdrk">&nbsp;</div> <ul class="list-set list-three-col"> <li><iframe src="https://www.youtube.com/embed/q0T_Af7khHw?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" height="151px" width="270px" _fsrb="true"></iframe> <h3>Reliable Quantification of Western Blot Data</h3> <p>Based on the article "A Defined Methodology for Reliable Quantification of Western Blot Data". Dr. S.C Taylor provides a direct comparison between results generated from camera-based imaging technologies and film using identical blots. Learn common pitfalls associated with film use and densitometric analysis.</p> <p class="caption">This video is based on the article published by S.C. Taylor et al. entitled <a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3840294/" target="_blank" rel="noopener noreferrer">"A Defined Methodology for Reliable Quantification of Western Blot Data"</a> (DOI 10.1007/s12033-013-9672-6).</p> </li> <li><iframe src="https://www.youtube.com/embed/iCgmeufGM2g?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" height="151px" width="270px" _fsrb="true"></iframe> <h3>Designing a Quantitative Western Blot Experiment to Avoid Common Pitfalls</h3> <p>The quantitative western blot methodology is described with a focus on solid experimental design and data analysis to achieve reproducible, publication quality results in the shortest time frame.</p> <p class="caption">This video is based on the article published by Drs. Taylor and Posch entitled: <a href="http://dx.doi.org/10.1155/2014/361590" target="_blank" rel="noopener noreferrer">"The Design of a Quantitative Western Blot Experiment"</a>.</p> </li> <li><iframe src="https://www.youtube.com/embed/QXg0r_DVlAs?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" height="151px" width="270px" _fsrb="true"></iframe> <h3>Quantitative Western Blotting: Optimization is the Key</h3> <p>Dr. S.C. Taylor details the common errors associated with performing quantitative western blots and how they can be addressed through a rigorous optimization methodology. Each step of the quantitative western blot workflow is discussed with a focus on optimization to assure quantitative, reproducible final data that reflect the experimental conditions.</p> </li> </ul> <img class="opt-horizdivider margin-bottom20" src="/webroot/web/images/opt/digital-pcr/horiz-gradient-divider.png" /> <iframe src="https://www.youtube.com/embed/NPeQ5TZ2qDk?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" height="245px" width="437px" _fsrb="true" class="right"></iframe> <h3 class="opt-subhead">Testimonial</h3> <p>Leukemia researcher Ernesto Diaz-Flores (University of California, San Francisco) describes how the ChemiDoc Imaging System helped him improve his western blot imaging, enabling him to identify relevant therapeutic targets. An avid photographer, Prof. Diaz-Flores also talks about the links between his art and his science and technologies that help him in both fields.</p> <p><a href="https://www.bioradiations.com/telling-the-story-of-three-western-blots-with-the-chemidoc-touch-imaging-system/" target="_blank" rel="noopener noreferrer"><strong>Read Full Article</strong></a></p> <img class="opt-horizdivider top-margin" src="/webroot/web/images/opt/digital-pcr/horiz-gradient-divider.png" /> <div class="opt-module-footer footer-twas-icon"><center> <ul class="no-ul-style mid-align"> <li> <div><img src="/webroot/web/images/general/global/english/have-questions-icon.png" border="0" alt="Consult with an Specialist" class="have-questions-icon" /></div> <div> <p class="in-line-message">Get answers to your questions about imaging</p> </div> <div><a href="https://info.bio-rad.com/ww-image-lab-twas.html" class="bttn-block no-target-style" target="_blank" rel="noopener noreferrer"><span class="bttn-orange">Consult with a Specialist</span></a></div> </li> </ul> </center></div> </div> <div><a name="resources"></a></div> <div class="opt-module optresources-bg"> <h2 class="nextp">Resources</h2> <div class="corner-box opt-western-blot-webinar linkpointer-blank"> <h2 class="nextp"><span class="featuredtxt">FEATURED WEBINAR</span>Understand the Factors That are Crucial to Successful Western Blotting</h2> <img src="/webroot/web/images/lsr/products/imaging_bioinformatics/category_feature/global/kenneth-oh-th.jpg" alt="Successful Western Blotting" class="left" /> <p>Join Bio-Rad western blotting expert Kenneth Oh and his guest speakers for a webinar series that explores the many factors that go into the design and execution of successful and repeatable western blots. Chemiluminescence or fluorescence, qualitative or quantitative blot, not only will we discuss the how-to's, we'll take a deeper dive and discuss the why's.</p> <p><a class="bttn-block no-target-style" onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Featured Webinar&amp;tgp_pcp_link=View Webinar','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="https://info.bio-rad.com/ww-PQDwebinars.html" target="_blank" rel="noopener noreferrer"><span class="bttn-orange">View Webinar</span></a></p> </div> <br /> <ul class="list-set list-two-col list-set-th"> <li><a href="/en-us/category/image-lab-touch-resources?ID=PJWAQ6E08O1Y "><img src="/webroot/web/images/lsr/products/imaging_bioinformatics/category_feature/global/image-lab-touch-video-th.gif" alt="Image Lab Touch Software Tutorials" /></a> <h3><a href="/en-us/category/image-lab-touch-resources?ID=PJWAQ6E08O1Y ">Image Lab Touch Software Resources</a></h3> <p>Bio-Rad provides instructional videos for Image Lab Touch Software 2.4, covering the primary applications of the ChemiDoc MP Imaging System. Topics include, logging in, Smart Tray Technology, capturing your image, and exporting data.</p> </li> <li><a href="/en-us/category/image-lab-software-resources?ID=PJWA0VTU86LJ"><img src="/webroot/web/images/lsr/products/imaging_bioinformatics/category_feature/global/image-lab-video-th.gif" alt="Image Lab Software Tutorials" /></a> <h3><a href="/en-us/category/image-lab-software-resources?ID=PJWA0VTU86LJ">Image Lab Software Resources</a></h3> <p>Bio-Rad provides tutorials for basic acquisition and advanced analysis use of Image Lab Software 6.0.1. Topics include, densitometric analysis, molecular weight, normalization, purity assessment, and more!</p> </li> </ul> <div class="dotteddivider dotteddividerdrk dotteddivider-less-top">&nbsp;</div> <h3 class="opt-subhead30">Application Guides</h3> <ul class="list-set list-three-col"> <li><a class="linkgeneration" onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Resources&amp;tgp_pcp_link=Western Blot Dr Troubleshooting Guide Img','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/en-us/applications-technologies/troubleshooting-western-blots-with-western-blot-doctor"><img src="/webroot/web/images/lsr/products/electrophoresis/category_feature/global/western-blot-guides-dr-troubleshooting.jpg" alt="Western Blot Dr. Troubleshooting Guide" /></a> <h3><a class="linkgeneration" onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Resources&amp;tgp_pcp_link=Western Blot Dr Troubleshooting Guide Txt','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/en-us/applications-technologies/troubleshooting-western-blots-with-western-blot-doctor">Western Blot Doctor Troubleshooting Guide</a></h3> <p>Our self-help troubleshooting guide covers solutions to many common and not-so-common western blotting issues and helps your blots look <span class="forcebreaks">their best.</span></p> </li> <li><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Resources&amp;tgp_pcp_link=Protein Blotting Guide Img','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/webroot/web/pdf/lsr/literature/Bulletin_2895.pdf" target="_blank" rel="noopener noreferrer" class="no-target-style"><img src="/webroot/web/images/lsr/products/electrophoresis/category_feature/global/western-blot-guides-blotting-guide.jpg" alt="Protein Blotting Guide" /></a> <h3><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Resources&amp;tgp_pcp_link=Protein Blotting Guide Txt','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/webroot/web/pdf/lsr/literature/Bulletin_2895.pdf" target="_blank" rel="noopener noreferrer" class="no-target-style">Protein Blotting Guide</a></h3> <p><span class="filesize">(PDF 7.9 MB)</span><br />Details on blotting technology, available products, and methods, plus tips, techniques, and troubleshooting.</p> </li> <li><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Resources&amp;tgp_pcp_link=Electrophoresis Guide Img','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/webroot/web/pdf/lsr/literature/Bulletin_6040.pdf" target="_blank" rel="noopener noreferrer" class="no-target-style"><img src="/webroot/web/images/lsr/products/electrophoresis/category_feature/global/western-blot-guides-electrophoresis-guide.jpg" alt="Electrophoresis Guide" /></a> <h3><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Resources&amp;tgp_pcp_link=Electrophoresis Guide Txt','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/webroot/web/pdf/lsr/literature/Bulletin_6040.pdf" target="_blank" rel="noopener noreferrer" class="no-target-style">Electrophoresis Guide</a></h3> <p><span class="filesize">(PDF 6.4 MB)</span><br />A guide to polyacrylamide gel electrophoresis and protein detection, including theory, product selection, protocols, and more.</p> </li> </ul> <div class="dotteddivider dotteddividerdrk dotteddivider-less-top">&nbsp;</div> <h3 class="opt-subhead30">Documents Library</h3> <ul class="list-set list-two-col list-set-th"> <li><a href="/webroot/web/pdf/lsr/literature/Bulletin_6873.pdf" target="_blank" rel="noopener noreferrer" class="no-target-style"><img src="/webroot/web/images/lsr/products/electrophoresis/category_feature/global/MA17-0012_S3_CV1_T2_01.png" /></a> <h3><a href="/webroot/web/pdf/lsr/literature/Bulletin_6873.pdf" target="_blank" rel="noopener noreferrer" class="no-target-style">ChemiDoc MP System Brochure</a><br /><span class="filesize">(PDF 1.26 MB)</span></h3> <p>Explore the features and benefits of the new high-end imaging system for the best fluorescence and chemiluminescence detection.</p> </li> <li><a href="/webroot/web/pdf/lsr/literature/Bulletin_RP0051.pdf" target="_blank" rel="noopener noreferrer" class="no-target-style"><img src="/webroot/web/images/lsr/products/electrophoresis/category_feature/global/MA17-0012_S3_CV1_T1_04.png" /> </a> <h3><a href="/webroot/web/pdf/lsr/literature/Bulletin_RP0051.pdf" target="_blank" rel="noopener noreferrer" class="no-target-style">Stain-Free Approach to Western Blotting</a><br /><span class="filesize">(PDF 864 KB)</span></h3> <p>Learn how to use stain-free technology for total protein normalization as an alternative to the standard blot normalization process.</p> </li> <li><a href="http://www.bio-rad.com/webroot/web/pdf/lsr/literature/Bulletin_6427.pdf" target="_blank" rel="noopener noreferrer" class="no-target-style"><img src="/webroot/web/images/lsr/products/electrophoresis/category_feature/global/MA17-0012_S3_CV1_T1_01.png" /></a> <h3><a href="http://www.bio-rad.com/webroot/web/pdf/lsr/literature/Bulletin_6427.pdf" target="_blank" rel="noopener noreferrer" class="no-target-style">V3 Western Workflow Brochure</a><br /><span class="filesize">(PDF 593 KB)</span></h3> <p>See how the V3 Western Workflow compares to traditional western blotting workflows.</p> </li> <li><a href="/webroot/web/pdf/lsr/literature/Bulletin_6363.pdf" target="_blank" class="no-target-style" rel="noopener noreferrer"><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/imaging_and_analysis/technology_detail/6363-pdf-th.png" alt="" /></a> <h3><a href="/webroot/web/pdf/lsr/literature/Bulletin_6363.pdf" target="_blank" rel="noopener noreferrer">Determining the Appropriate Sample Load When Using a Stain-Free V3 Western Workflow</a><br /><span class="filesize">(182 KB)</span></h3> </li> <li><a href="/webroot/web/pdf/lsr/literature/Bulletin_6366.pdf" target="_blank" class="no-target-style" rel="noopener noreferrer"><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/imaging_and_analysis/technology_detail/6366-pdf-th.png" alt="" /></a> <h3><a href="/webroot/web/pdf/lsr/literature/Bulletin_6366.pdf" target="_blank" rel="noopener noreferrer">Validating the Expression Consistency of a Housekeeping Protein</a><br /><span class="filesize">(198 KB)</span></h3> </li> </ul> <div class="dotteddivider dotteddividerdrk dotteddivider-less-top">&nbsp;</div> <h3 class="opt-subhead30">Publications</h3> <ul class="list-set list-two-col"> <li> <h3><a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3840294/" target="_blank" rel="noopener noreferrer">A Defined Methodology for Reliable Quantification of Western Blot Data</a></h3> <p>Learn about the methodology to obtain reliable quantitative data from chemiluminescent western blots using standardization procedures coupled with the updated reagents and detection methods.</p> </li> <li> <h3><a href="https://www.hindawi.com/journals/bmri/2014/361590/" target="_blank" rel="noopener noreferrer">The Design of a Quantitative Western Blot Experiment</a></h3> <p>Read a summary about a complete western blot workflow with a focus on sample preparation and data analysis for quantitative western blotting.</p> </li> </ul> </div> <div><a name="related"></a></div> <div class="opt-module"> <h2>Related Products</h2> <ul class="list-set list-two-col list-set-th"> <li><a href="/en-us/product/mini-protean-precast-gels/mini-protean-tgx-stain-free-precast-gels?tab=ordering"><img src="/webroot/web/images/lsr/products/electrophoresis/category_thumb/global/protean-tgx-stain-free-precast-gels-thumb-mini-protean-precast-gels.jpg" style="width: 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Turbo System is a rapid protein transfer apparatus that can transfer protein to membrane in as little as 3 minutes. Trans-Blot Turbo Transfer Packs provide greater transfer efficiency in less time.</p> </li> <li><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Related Products&amp;tgp_pcp_link=Protein Ladders and Standards (Markers)','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="../category/protein-ladders-standards-markers"><img src="/webroot/web/images/lsr/products/imaging_bioinformatics/category_feature/global/protein-ladders-standards-related-prdcts-th.jpg" alt="Protein Ladders and Standards (Markers)" /></a> <h3><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Related Products&amp;tgp_pcp_link=Protein Ladders and Standards (Markers)','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="../category/protein-ladders-standards-markers">Protein Ladders and Standards (Markers)</a></h3> <p>Prestained and unstained molecular weight standards for protein electrophoresis applications including SDS-PAGE, western blotting, 2-D PAGE, and isoelectric focusing (IEF).</p> </li> <li><a href="/en-us/product/clarity-clarity-max-western-ecl-blotting-substrates?ID=e08909eb-3369-42ec-a585-df189004268f"><img src="/webroot/web/images/lsr/products/electrophoresis/product_thumb/global/immunodetection-reagents-and-kits-thumb-clarity-western-ecl-substrate-feature.jpg" alt="" /></a> <h3><a href="/en-us/product/clarity-clarity-max-western-ecl-blotting-substrates?ID=e08909eb-3369-42ec-a585-df189004268f">Clarity ECL </a></h3> <p>Clarity family of Western ECL Substrates provides simple, high-performance solutions for all your western chemiluminescence needs.&nbsp; &nbsp;&nbsp;</p> </li> <li><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Related Products&amp;tgp_pcp_link=SureBeads Protein G Magnetic Beads','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/en-de/product/surebeads-protein-g-magnetic-beads?ID=N6AQ2QE8Z"><img border="0" alt="" src="/webroot/web/images/lsr/products/protein_sample_preparation/product_thumb/global/protein-g-magnetic-beads-thumb.jpg" /></a> <h3><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Related Products&amp;tgp_pcp_link=SureBeads Protein G Magnetic Beads','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/en-de/product/surebeads-protein-g-magnetic-beads?ID=N6AQ2QE8Z">SureBeads Protein G Magnetic Beads</a></h3> <p>SureBeads Protein G Magnetic Beads enable fast, easy, consistent immunoprecipitation without centrifugation.</p> </li> </ul> <p><a class="scrollup" href="#">Scroll</a> <script src="/webroot/web/js/protein-marker-min.js" type="text/javascript"></script> <script type="text/javascript" src="/webroot/web/js/apptech-opt-conveter-min.js"></script> </p> </div> <script type="text/javascript">// <![CDATA[ $(document).ready(function () { // START Ribbon $( '.ribbonThird' ).hover( function() { $( '.ribbonThird:first' ).removeClass( 'active' ); $(this).addClass( 'active' ); }, function() { $(this).removeClass( 'active' ); }); // START Ribbon }); // ]]></script> <script type="text/javascript" src="/webroot/web/js/apptech-opt-conveter-min.js"></script> Anton Ross Compare X-Ray Film to Digital Imaging Systems | Bio-Rad.com Bio-Rad's digital imaging systems provide accurate quantitation of a range of signal intensities in western blotting when compared to X-Ray film. x-ray film, digital imaging systems, imaging systems, comparing x-ray to film, compare x-ray to film, western blotting, quantitation, x-ray versus digital imaging, x-ray vs digital imaging 02/26/19 11:17 PM 08/22/29 09:05 AM AR,BO,BR,CA,CL,CO,CR,DO,EC,SV,GU,GT,HT,HN,JM,MX,NI,PA,PY,PE,PR,TT,US,UY,VE,AU,BD,BN,KH,CN,HK,IN,ID,JP,KR,MO,MY,FM,NP,NZ,PW,PG,PH,SG,SB,LK,TW,TH,TO,VU,AL,DZ,AI,AM,AT,AZ,BH,BE,BA,BW,BG,BF,CM,HR,CY,CZ,DK,EG,ER,EE,ET,FO,FI,FR,GF,PF,GA,GE,DE,GH,GR,GP,HU,IS,IE,IL,IT,JO,KZ,KE,XK,KW,LV,LB,LI,LT,LU,MK,MG,ML,MT,MQ,MU,MD,MS,MA,NL,NG,NO,OM,PK,PS,PL,PT,QA,RO,RU,ST,SA,SN,RS,SK,SI,ZA,ES,SE,CH,TZ,TG,TN,TR,UG,UA,AE,UK,UZ,VA,EH,YE,VN LSR /LSR/Technologies/Imaging_and_Analysis N 0 <div class="pagenav"><img src="/webroot/web/images/opt/prodnav-arrow-top.png" alt="" /> <table border="0"> <tbody> <tr> <td class="pagenavhead"> <h3>On This Page</h3> </td> <td class="pagenavfirst"><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Page Nav&amp;tgp_pcp_link=Advantages of Digital Imaging Systems','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="#advantages">Advantages of Digital Imaging Systems</a></td> <td><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Page Nav&amp;tgp_pcp_link=ChemiDoc Imaging System','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="#ChIS">ChemiDoc Imaging System</a></td> <td><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Page Nav&amp;tgp_pcp_link=Publication Requirements','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="#publication">Publication Requirements</a></td> <td><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Page Nav&amp;tgp_pcp_link=Designing Experiments','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="#designing">Designing Experiments</a></td> <td><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Page Nav&amp;tgp_pcp_link=Resources','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="#resources">Resources</a></td> <td class="pagenavlast"><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Page Nav&amp;tgp_pcp_link=Related Products','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="#related">Related Products</a></td> </tr> </tbody> </table> </div> <div class="opt-module opt-module-first opt-module-noborder"> <div class="corner-box corner-box-promo linkpointer-blank"><img src="/webroot/web/images/general/global/english/have-questions-icon.png" border="0" alt="Consult with an Specialist" style="width: 50px; height: auto;" /> <p><strong>Need help with imaging?</strong></p> <a href="https://info.bio-rad.com/ww-image-lab-twas.html" class="bttn-block no-target-style" target="_blank" rel="noopener noreferrer"><span class="bttn-orange">Consult with a Specialist</span></a></div> <p>X-ray film has long been considered the gold standard of sensitivity in western blotting, but the broad dynamic range enabled by digital imaging systems allows for accurate quantitation of a range of signal intensities. With recent technological advances, can researchers finally have the best of both worlds?</p> </div> <div><a name="advantages"></a></div> <div class="opt-module bg-f0f9f8 opt-module-noborder"> <h2 class="nextp">Advantages of Digital Imaging Systems</h2> <iframe src="https://www.youtube.com/embed/pzIRIrhgG2M?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" height="245px" width="437px" _fsrb="true" class="right"></iframe> <p><strong>Key benefits to using this technology include:</strong></p> <ul> <li>Wide dynamic range = &gt; 4 logs</li> <li>Quantitative</li> <li>Digital documentation</li> <li>Environmentally friendly &mdash; no chemicals!</li> <li>No costly consumables (film)</li> <li>Initial capital investment typically recovered <br />in as little as 1 year</li> </ul> <div class="corner-box bg-white top-margin"> <h2 class="no-bttm-pddng">See the Data</h2> <h3 class="opt-subhead30">The sensitivity of film, with a broad dynamic range and superior signal-to-noise.</h3> <center><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/imaging_and_analysis/technology_detail/low-signal-detection-film-data-sensitivity.png" alt="" class="top-margin" /></center> <p class="caption topmargin-1em"><strong>Comparison of sensitivity between the ChemiDoc MP Imaging System and film. A.</strong> Western blot analysis of LacI expression was conducted using 2x serial dilutions (starting at 0.31 &mu;g protein) of E. coli cell lysate. The membranes were either imaged on the ChemiDoc MP Imaging System for 30 sec or exposed to film for 30 sec. <strong>B.</strong> Band intensities, normalized to lane 7 background, illustrate the ability of the ChemiDoc MP Imaging System to detect low signal bands at the same exposure time as film. The red boxes represent the limited linear dynamic range of film. ChemiDoc MP Imaging System, 30 sec ( <span class="bar-graph-key bg-95cb45">&nbsp;</span> ); film, 30 sec ( <span class="bar-graph-key bg-184a74">&nbsp;</span> ).</p> <center><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/imaging_and_analysis/technology_detail/reveal-faint-bands-film-data-sensitivity.png" alt="" class="top-margin" /></center> <p class="caption topmargin-1em"><strong>Side-by-side comparison between the ChemiDoc Imaging System and film.</strong> Levels of the three isoforms of the pro-apoptotic protein Bim were measured in various cell lines using western blot analysis. The membranes were either imaged on the ChemiDoc Imaging System for 30 sec or exposed to film for 30 sec to compare detection sensitivities. As shown by the overlay graph, the ChemiDoc Imaging System was better able to detect faint protein bands than film.</p> </div> &nbsp;<br clear="all" /> <iframe src="https://www.youtube.com/embed/xAUFWRONV_U?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" mce_src="https://www.youtube.com/embed/nVsLxeT8Reg?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" height="245px" width="437px" _fsrb="true" class="left"></iframe> <h3 class="opt-subhead">ChemiDoc MP Imaging System: Imagine No Compromises</h3> <p>With the ChemiDoc MP Imaging System, the superior quantitation of digital imaging meets the sensitivity of film. This best-in-class system images and analyzes gels and western blots. It is designed for multiplex fluorescent western blotting, chemiluminescence detection, general gel documentation, and stain-free technology imaging needs.</p> <p><a class="bttn-block no-target-style" onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Product Spotlight&amp;tgp_pcp_link=Learn More','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/en-us/category/imaging-systems-software?ID=f97d1fa1-16ff-4c73-9eae-ef74be3a3f95"><span class="bttn-orange">Learn More</span></a></p> </div> <div><a name="ChIS"></a></div> <div class="opt-module opt-module-noborder bg-f9f9e4"> <h2>ChemiDoc Imaging System</h2> <h3 class="opt-subhead30 top-margin-neg">Better Options. Better Results.</h3> <div id="interactiveRibbonOuter"> <div class="ribbonThird" style="background-image: url('/webroot/web/images/lsr/global/english/solutions/technologies/Fluorescent-Western-Blotting/stain-free-square.png'); background-repeat: no-repeat;"> <div class="ribbonThirdInner"> <div class="ribbonHeader"> <div class="ribbon2"> <div class="ribbon2Center"> <h4 class="gy05">Stain-Free<br /><small>Total Protein Normalization</small></h4> </div> </div> <div class="ribbon2Right"></div> </div> <div class="ribbonDesc"> <p>Learn how to properly design a quantitative western blotting experiment and faithfully compare protein expression levels, without concern for changing or overexpressed HKP&rsquo;s. <br /><a href="/en-sg/applications-technologies/total-protein-normalization?ID=PODYJQRT8IG9">See Data &raquo;</a></p> </div> </div> </div> <div class="ribbonThird" style="background-image: url('/webroot/web/images/lsr/global/english/solutions/technologies/Fluorescent-Western-Blotting/chemiluminescence.jpg'); background-repeat: no-repeat;"> <div class="ribbonThirdInner"> <div class="ribbonHeader"> <div class="ribbon2"> <div class="ribbon2Center"> <h4 class="gy05">Chemiluminescence<br /><small>Replaces Film</small></h4> </div> </div> <div class="ribbon2Right"></div> </div> <div class="ribbonDesc"> <p>Save money on consumables and reduce environmental waste without compromising on performance. Get the sensitivity of film and the convenience of digital documentation. <br /><a href="/en-sg/applications-technologies/comparing-x-ray-film-digital-imaging-systems?ID=PNK1ECTU86LJ">See Data &raquo;</a></p> </div> </div> </div> <div class="ribbonThird" style="background-image: url('/webroot/web/images/lsr/global/english/solutions/technologies/Fluorescent-Western-Blotting/bio-rad-fluorescence.jpg'); background-repeat: no-repeat;"> <div class="ribbonThirdInner"> <div class="ribbonHeader"> <div class="ribbon2"> <div class="ribbon2Center"> <h4 class="gy05">Multiplex Fluorescence<br /><small>3 Colors</small></h4> </div> </div> <div class="ribbon2Right"></div> </div> <div class="ribbonDesc"> <p>Save sample, avoid the errors commonly associated with stripping, reprobing, and cutting blots and answer more complex biological questions by optimizing your experiments to fluorescence. <br /><a href="/en-sg/applications-technologies/fluorescent-western-blotting?ID=PJW4ANE08O1Y">See Data &raquo;</a></p> </div> </div> </div> </div> </div> <div><a name="publication"></a></div> <div class="opt-module opt-module-noborder"> <h2 class="nextp">Publication Requirements</h2> <h3 class="opt-subhead">Need another reason to get out of the darkroom?</h3> <p>Several journals have recently revised their guidelines for publication, providing guidance around reproducibility and quantitation.</p> <ul class="list-set list-four-col topmargin-1em"> <li><center><a href="https://www.nature.com/authors/policies/image.html" target="_blank" class="no-target-class" rel="noopener noreferrer"><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/imaging_and_analysis/technology_detail/website-nature-4col-th.png" alt="" /></a> <h3><a href="https://www.nature.com/authors/policies/image.html" target="_blank" class="no-target-class" rel="noopener noreferrer">Nature</a></h3> </center></li> <li><center><a href="https://science.sciencemag.org/content/346/6210/679.full" target="_blank" class="no-target-class" rel="noopener noreferrer"><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/imaging_and_analysis/technology_detail/website-science-4col-th.png" alt="" /></a> <h3><a href="https://science.sciencemag.org/content/346/6210/679.full" target="_blank" rel="noopener noreferrer">Science</a></h3> </center></li> <li><center><a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4705984/" target="_blank" class="no-target-class" rel="noopener noreferrer"><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/imaging_and_analysis/technology_detail/website-ncbi-4col-th.png" alt="" /></a> <h3><a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4705984/" target="_blank" rel="noopener noreferrer">National Center for Biotechnology Information</a></h3> </center></li> <li><center><a href="https://www.jbc.org/site/misc/ifora.xhtml" target="_blank" class="no-target-class" rel="noopener noreferrer"><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/imaging_and_analysis/technology_detail/website-jbc-4col-th.png" alt="" /></a> <h3><a href="https://www.jbc.org/site/misc/ifora.xhtml" target="_blank" rel="noopener noreferrer">Journal of Biological Chemistry</a></h3> </center></li> </ul> <div class="corner-box corner-box-tpn linkpointer-blank"> <h3><a href="https://www.bioradiations.com/defining-the-new-normal-in-quantitative-western-blot-data" target="_blank" rel="noopener noreferrer">Defining the New Normal in Quantitative Western Blot Data</a></h3> <p>Learn why total protein normalization (TPN) is a superior method for quantitative western blotting analysis.</p> </div> <h3 class="opt-subhead30 top-margin">Publication requirements are changing. Are your blots ready?</h3> <ul class="flex-container flex-3-col flex-col-stroke leaf-border flex-pub-req"> <li> <h3><span>1</span></h3> <p><strong>"House-keeping proteins should not be used for normalization without evidence that experimental manipulations do not affect their expression"*</strong></p> <p><strong>Solution:</strong> Use Total Protein Normalization (TPN) instead of house-keeping proteins</p> <ul class="no-list-pddng disc-list"> <li>Bio-Rad's stain-free workflow makes TPN easy! This technology is proven with over 100 publications, allows for improved accuracy and will make your data more reliable</li> </ul> </li> <li> <h3><span>2</span></h3> <p><strong>"Methods including detection of enhanced chemiluminescence using X-ray film have a very limited dynamic range"*</strong></p> <p><strong>Solution:</strong> Use an imaging system with up to 4 logs of dynamic range</p> <ul class="no-list-pddng disc-list"> <li>Bio-Rad's ChemiDoc System &mdash; an imager with the sensitivity of film</li> <li>Simple image generation and quantitation using Image Lab Software</li> </ul> </li> <li> <h3><span>3</span></h3> <p><strong>"A description of the data supporting the specificity of all antibodies is required"*</strong></p> <p><strong>Solution:</strong> Fully validated antibodies</p> <ul class="no-list-pddng disc-list"> <li>Bio-Rad's PrecisionAb Antibodies offer: documented specificity and validation protocols</li> </ul> </li> </ul> <p class="caption top-margin">* Revised guidelines for authors from <a href="http://www.jbc.org/site/misc/ifora.xhtml#blots" target="_blank" rel="noopener noreferrer">The Journal of Biological Chemistry</a></p> <div class="dotteddivider dotteddividerdrk dotteddivider-less-top">&nbsp;</div> <h3 class="opt-subhead">Total Protein Normalization provides accurate quantitation of target proteins.</h3> <p>Normalizing blots with housekeeping proteins (HKPs) requires time, money, and additional sample to validate HKP expression and linearity. Total protein normalization using stain-free technology and the ChemiDoc or ChemiDoc MP System allows normalization across a wide dynamic range. Get better, more publishable data without lengthy optimization.</p> <img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/imaging_and_analysis/technology_detail/acurate-tpn.png" class="topmargin-1em" alt="" /> <p class="caption topmargin-1em"><strong>Linearity comparison of stain-free total protein measurement and immunodetection of three housekeeping proteins in 10&ndash;50 &micro;g of HeLa cell lysate.</strong></p> <div class="corner-box bg-white no-bttm-pddng topmargin-1em corner-box-stainfree"> <h3 class="opt-subhead30">Learn more about Stain-free Technology</h3> <ul class="list-set list-two-col list-set-th"> <li><a href="https://www.bioradiations.com/trends-in-protein-separation-and-analysis-the-advance-of-stain-free-technology/" target="_blank" rel="noopener noreferrer"><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/imaging_and_analysis/technology_detail/" alt="" /></a> <h3><a href="https://www.bioradiations.com/trends-in-protein-separation-and-analysis-the-advance-of-stain-free-technology/" target="_blank" rel="noopener noreferrer">Trends in Protein Separation and Analysis &mdash; the Advance of Stain-Free Technology</a></h3> <p>This article focuses on the theory and concepts surrounding stain-free technology as well as other traditional methods of protein staining.</p> </li> <li> <h3><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=AppTech&amp;tgp_pcp_link=Stain-Free Imaging Technology','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/en-us/applications-technologies/stain-free-imaging-technology">Stain-Free Imaging Technology</a></h3> <p>Visualize proteins on gels and blots without staining and destaining steps, and use total protein normalization for quantitative western blots.</p> </li> </ul> </div> <h3 class="opt-subhead30">Bio-Rad's V3 Western Workflow Incorporates TPN for Normalization of Western Blot Data</h3> <iframe src="https://www.youtube.com/embed/RWsDtTqDW_o?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" height="245px" width="437px" class="right"></iframe> <p>V3 Western Workflow includes:</p> <ul> <li>Stain-free visualization of total proteins on gel after electrophoresis</li> <li>Quantification of total proteins in a lane after transfer</li> <li>Normalization of bands to total proteins by specialized software</li> </ul> <p>Find out why these 3 steps are crucial to generating quantitative western blot data.</p> <p><a class="bttn-block linkgeneration" onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=V3&amp;tgp_pcp_link=Learn more','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/en-us/product/v3-western-workflow"><span class="bttn-orange">Learn More</span></a> &nbsp;<a class="bttn-block no-target-style" onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=V3&amp;tgp_pcp_link=Request a Demo','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="https://info.bio-rad.com/ww-V3-Central-Umbrella-btwas.html" target="_blank" rel="noopener noreferrer"><span class="bttn-green">Request a Demo</span></a></p> </div> <div class="opt-module white opt-module-precisionab-feature linkpointer"> <h2 class="nextp white"><span class="featuredtxt">FEATURED PRODUCT</span>PrecisionAb Validated Primary Antibodies<br />for Western Blotting</h2> <ul class="list-orange-arrow"> <li class="white">Comprehensive selection of antibodies for cancer and neuroscience research</li> <li class="white">Highly specific and sensitive</li> <li class="white">Validated against up to 12 lysates</li> <li class="white">Stringent QC criteria for superior consistency</li> </ul> <p><a class="bttn-block" onmousedown="dcsMultiTrack('DCS.dcsqry','? tgp_pcp_link_group=PrecisionAb Validated Western Blotting Antibodies&amp;tgp_pcp_link=Learn more','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999'); return false;" href="/en-us/product/precisionab-validated-western-blotting-antibodies"><span class="bttn-orange">Find Your Antibody</span></a></p> </div> <div><a name="designing"></a></div> <div class="opt-module opt-module-noborder no-bttm-pddng"> <h2>Design your Western Blotting Experiments with Confidence</h2> <iframe src="https://www.youtube.com/embed/zrfAHAo4xT0?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" height="245px" width="437px" _fsrb="true" class="right"></iframe> <p>As an editor of several journals, Professor Aldrin V. Gomes routinely observes western blots that are not properly conducted. The generally low quality of western blotting data has led scientists to question whether we can trust protein quantification when using western blots. Reducing errors and misinterpretations requires standardized procedure and proper controls. Dr. Gomes discusses the following key factors for high-confidence western blot data:</p> <ol> <li>Quality of the primary and secondary antibodies and the blocking reagent</li> <li>Proper target protein controls</li> <li>Choice of total protein loading control vs. housekeeping protein loading control to normalize the target protein measurement</li> </ol> <p>Prof. Gomes also presents tips to reduce the overall time required for first-rate western blots, and how to minimize the pain of going through a long procedure.</p> <div class="dotteddivider dotteddividerdrk">&nbsp;</div> <ul class="list-set list-three-col"> <li><iframe src="https://www.youtube.com/embed/q0T_Af7khHw?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" height="151px" width="270px" _fsrb="true"></iframe> <h3>Reliable Quantification of Western Blot Data</h3> <p>Based on the article "A Defined Methodology for Reliable Quantification of Western Blot Data". Dr. S.C Taylor provides a direct comparison between results generated from camera-based imaging technologies and film using identical blots. Learn common pitfalls associated with film use and densitometric analysis.</p> <p class="caption">This video is based on the article published by S.C. Taylor et al. entitled <a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3840294/" target="_blank" rel="noopener noreferrer">"A Defined Methodology for Reliable Quantification of Western Blot Data"</a> (DOI 10.1007/s12033-013-9672-6).</p> </li> <li><iframe src="https://www.youtube.com/embed/iCgmeufGM2g?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" height="151px" width="270px" _fsrb="true"></iframe> <h3>Designing a Quantitative Western Blot Experiment to Avoid Common Pitfalls</h3> <p>The quantitative western blot methodology is described with a focus on solid experimental design and data analysis to achieve reproducible, publication quality results in the shortest time frame.</p> <p class="caption">This video is based on the article published by Drs. Taylor and Posch entitled: <a href="http://dx.doi.org/10.1155/2014/361590" target="_blank" rel="noopener noreferrer">"The Design of a Quantitative Western Blot Experiment"</a>.</p> </li> <li><iframe src="https://www.youtube.com/embed/QXg0r_DVlAs?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" height="151px" width="270px" _fsrb="true"></iframe> <h3>Quantitative Western Blotting: Optimization is the Key</h3> <p>Dr. S.C. Taylor details the common errors associated with performing quantitative western blots and how they can be addressed through a rigorous optimization methodology. Each step of the quantitative western blot workflow is discussed with a focus on optimization to assure quantitative, reproducible final data that reflect the experimental conditions.</p> </li> </ul> <img class="opt-horizdivider margin-bottom20" src="/webroot/web/images/opt/digital-pcr/horiz-gradient-divider.png" /> <iframe src="https://www.youtube.com/embed/NPeQ5TZ2qDk?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" height="245px" width="437px" _fsrb="true" class="right"></iframe> <h3 class="opt-subhead">Testimonial</h3> <p>Leukemia researcher Ernesto Diaz-Flores (University of California, San Francisco) describes how the ChemiDoc Imaging System helped him improve his western blot imaging, enabling him to identify relevant therapeutic targets. An avid photographer, Prof. Diaz-Flores also talks about the links between his art and his science and technologies that help him in both fields.</p> <p><a href="https://www.bioradiations.com/telling-the-story-of-three-western-blots-with-the-chemidoc-touch-imaging-system/" target="_blank" rel="noopener noreferrer"><strong>Read Full Article</strong></a></p> <img class="opt-horizdivider top-margin" src="/webroot/web/images/opt/digital-pcr/horiz-gradient-divider.png" /> <div class="opt-module-footer footer-twas-icon"><center> <ul class="no-ul-style mid-align"> <li> <div><img src="/webroot/web/images/general/global/english/have-questions-icon.png" border="0" alt="Consult with an Specialist" class="have-questions-icon" /></div> <div> <p class="in-line-message">Get answers to your questions about imaging</p> </div> <div><a href="https://info.bio-rad.com/ww-image-lab-twas.html" class="bttn-block no-target-style" target="_blank" rel="noopener noreferrer"><span class="bttn-orange">Consult with a Specialist</span></a></div> </li> </ul> </center></div> </div> <div><a name="resources"></a></div> <div class="opt-module optresources-bg"> <h2 class="nextp">Resources</h2> <div class="corner-box opt-western-blot-webinar linkpointer-blank"> <h2 class="nextp"><span class="featuredtxt">FEATURED WEBINAR</span>Understand the Factors That are Crucial to Successful Western Blotting</h2> <img src="/webroot/web/images/lsr/products/imaging_bioinformatics/category_feature/global/kenneth-oh-th.jpg" alt="Successful Western Blotting" class="left" /> <p>Join Bio-Rad western blotting expert Kenneth Oh and his guest speakers for a webinar series that explores the many factors that go into the design and execution of successful and repeatable western blots. Chemiluminescence or fluorescence, qualitative or quantitative blot, not only will we discuss the how-to's, we'll take a deeper dive and discuss the why's.</p> <p><a class="bttn-block no-target-style" onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Featured Webinar&amp;tgp_pcp_link=View Webinar','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="https://info.bio-rad.com/ww-PQDwebinars.html" target="_blank" rel="noopener noreferrer"><span class="bttn-orange">View Webinar</span></a></p> </div> <br /> <ul class="list-set list-two-col list-set-th"> <li><a href="/en-us/category/image-lab-touch-resources?ID=PJWAQ6E08O1Y "><img src="/webroot/web/images/lsr/products/imaging_bioinformatics/category_feature/global/image-lab-touch-video-th.gif" alt="Image Lab Touch Software Tutorials" /></a> <h3><a href="/en-us/category/image-lab-touch-resources?ID=PJWAQ6E08O1Y ">Image Lab Touch Software Resources</a></h3> <p>Bio-Rad provides instructional videos for Image Lab Touch Software 2.4, covering the primary applications of the ChemiDoc MP Imaging System. Topics include, logging in, Smart Tray Technology, capturing your image, and exporting data.</p> </li> <li><a href="/en-us/category/image-lab-software-resources?ID=PJWA0VTU86LJ"><img src="/webroot/web/images/lsr/products/imaging_bioinformatics/category_feature/global/image-lab-video-th.gif" alt="Image Lab Software Tutorials" /></a> <h3><a href="/en-us/category/image-lab-software-resources?ID=PJWA0VTU86LJ">Image Lab Software Resources</a></h3> <p>Bio-Rad provides tutorials for basic acquisition and advanced analysis use of Image Lab Software 6.0.1. Topics include, densitometric analysis, molecular weight, normalization, purity assessment, and more!</p> </li> </ul> <div class="dotteddivider dotteddividerdrk dotteddivider-less-top">&nbsp;</div> <h3 class="opt-subhead30">Application Guides</h3> <ul class="list-set list-three-col"> <li><a class="linkgeneration" onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Resources&amp;tgp_pcp_link=Western Blot Dr Troubleshooting Guide Img','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/en-us/applications-technologies/troubleshooting-western-blots-with-western-blot-doctor"><img src="/webroot/web/images/lsr/products/electrophoresis/category_feature/global/western-blot-guides-dr-troubleshooting.jpg" alt="Western Blot Dr. Troubleshooting Guide" /></a> <h3><a class="linkgeneration" onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Resources&amp;tgp_pcp_link=Western Blot Dr Troubleshooting Guide Txt','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/en-us/applications-technologies/troubleshooting-western-blots-with-western-blot-doctor">Western Blot Doctor Troubleshooting Guide</a></h3> <p>Our self-help troubleshooting guide covers solutions to many common and not-so-common western blotting issues and helps your blots look <span class="forcebreaks">their best.</span></p> </li> <li><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Resources&amp;tgp_pcp_link=Protein Blotting Guide Img','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/webroot/web/pdf/lsr/literature/Bulletin_2895.pdf" target="_blank" rel="noopener noreferrer" class="no-target-style"><img src="/webroot/web/images/lsr/products/electrophoresis/category_feature/global/western-blot-guides-blotting-guide.jpg" alt="Protein Blotting Guide" /></a> <h3><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Resources&amp;tgp_pcp_link=Protein Blotting Guide Txt','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/webroot/web/pdf/lsr/literature/Bulletin_2895.pdf" target="_blank" rel="noopener noreferrer" class="no-target-style">Protein Blotting Guide</a></h3> <p><span class="filesize">(PDF 7.9 MB)</span><br />Details on blotting technology, available products, and methods, plus tips, techniques, and troubleshooting.</p> </li> <li><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Resources&amp;tgp_pcp_link=Electrophoresis Guide Img','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/webroot/web/pdf/lsr/literature/Bulletin_6040.pdf" target="_blank" rel="noopener noreferrer" class="no-target-style"><img src="/webroot/web/images/lsr/products/electrophoresis/category_feature/global/western-blot-guides-electrophoresis-guide.jpg" alt="Electrophoresis Guide" /></a> <h3><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Resources&amp;tgp_pcp_link=Electrophoresis Guide Txt','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/webroot/web/pdf/lsr/literature/Bulletin_6040.pdf" target="_blank" rel="noopener noreferrer" class="no-target-style">Electrophoresis Guide</a></h3> <p><span class="filesize">(PDF 6.4 MB)</span><br />A guide to polyacrylamide gel electrophoresis and protein detection, including theory, product selection, protocols, and more.</p> </li> </ul> <div class="dotteddivider dotteddividerdrk dotteddivider-less-top">&nbsp;</div> <h3 class="opt-subhead30">Documents Library</h3> <ul class="list-set list-two-col list-set-th"> <li><a href="/webroot/web/pdf/lsr/literature/Bulletin_6873.pdf" target="_blank" rel="noopener noreferrer" class="no-target-style"><img src="/webroot/web/images/lsr/products/electrophoresis/category_feature/global/MA17-0012_S3_CV1_T2_01.png" /></a> <h3><a href="/webroot/web/pdf/lsr/literature/Bulletin_6873.pdf" target="_blank" rel="noopener noreferrer" class="no-target-style">ChemiDoc MP System Brochure</a><br /><span class="filesize">(PDF 1.26 MB)</span></h3> <p>Explore the features and benefits of the new high-end imaging system for the best fluorescence and chemiluminescence detection.</p> </li> <li><a href="/webroot/web/pdf/lsr/literature/Bulletin_RP0051.pdf" target="_blank" rel="noopener noreferrer" class="no-target-style"><img src="/webroot/web/images/lsr/products/electrophoresis/category_feature/global/MA17-0012_S3_CV1_T1_04.png" /> </a> <h3><a href="/webroot/web/pdf/lsr/literature/Bulletin_RP0051.pdf" target="_blank" rel="noopener noreferrer" class="no-target-style">Stain-Free Approach to Western Blotting</a><br /><span class="filesize">(PDF 864 KB)</span></h3> <p>Learn how to use stain-free technology for total protein normalization as an alternative to the standard blot normalization process.</p> </li> <li><a href="http://www.bio-rad.com/webroot/web/pdf/lsr/literature/Bulletin_6427.pdf" target="_blank" rel="noopener noreferrer" class="no-target-style"><img src="/webroot/web/images/lsr/products/electrophoresis/category_feature/global/MA17-0012_S3_CV1_T1_01.png" /></a> <h3><a href="http://www.bio-rad.com/webroot/web/pdf/lsr/literature/Bulletin_6427.pdf" target="_blank" rel="noopener noreferrer" class="no-target-style">V3 Western Workflow Brochure</a><br /><span class="filesize">(PDF 593 KB)</span></h3> <p>See how the V3 Western Workflow compares to traditional western blotting workflows.</p> </li> <li><a href="/webroot/web/pdf/lsr/literature/Bulletin_6363.pdf" target="_blank" class="no-target-style" rel="noopener noreferrer"><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/imaging_and_analysis/technology_detail/6363-pdf-th.png" alt="" /></a> <h3><a href="/webroot/web/pdf/lsr/literature/Bulletin_6363.pdf" target="_blank" rel="noopener noreferrer">Determining the Appropriate Sample Load When Using a Stain-Free V3 Western Workflow</a><br /><span class="filesize">(182 KB)</span></h3> </li> <li><a href="/webroot/web/pdf/lsr/literature/Bulletin_6366.pdf" target="_blank" class="no-target-style" rel="noopener noreferrer"><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/imaging_and_analysis/technology_detail/6366-pdf-th.png" alt="" /></a> <h3><a href="/webroot/web/pdf/lsr/literature/Bulletin_6366.pdf" target="_blank" rel="noopener noreferrer">Validating the Expression Consistency of a Housekeeping Protein</a><br /><span class="filesize">(198 KB)</span></h3> </li> </ul> <div class="dotteddivider dotteddividerdrk dotteddivider-less-top">&nbsp;</div> <h3 class="opt-subhead30">Publications</h3> <ul class="list-set list-two-col"> <li> <h3><a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3840294/" target="_blank" rel="noopener noreferrer">A Defined Methodology for Reliable 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125px; height: auto;" /></a> <h3><a href="/en-us/product/mini-protean-precast-gels/mini-protean-tgx-stain-free-precast-gels?tab=ordering">Mini-PROTEAN TGX Stain-free Precast Gels</a></h3> <p>Mini-PROTEAN TGX Stain-Free Precast Gels for polyacrylamide gel electrophoresis (PAGE) allow for fast run times, high transfer efficiency, and rapid detection of proteins with stain-free enabled imagers.</p> </li> <li><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Related Products&amp;tgp_pcp_link=PrecisionAb Validated Western Blotting Antibodies','DCSext.tgp_pcp_link_group','','DCSext.tgp_pcp_link','','WT.dl','99999');" href="/en-us/product/precisionab-validated-western-blotting-antibodies"><img src="/webroot/web/images/lsr/products/bioplex_suspension_array/category_feature/global/precisionab-related-prod.jpg" alt="PrecisionAb Validated Western Blotting Antibodies" /></a> <h3><a onmousedown="dcsMultiTrack('DCS.dcsqry','?tgp_pcp_link_group=Related 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