Troubleshooting Western Blots with the Western Blot Doctor™

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en-us MIW4HR15 Western Blot Doctor Troubleshooting Western Blots with the Western Blot Doctor&#153; /webroot/web/html/lsr/solutions/technologies/western_blotting <script type="text/javascript"><!-- $(document).ready(function(){ $('.analysismethods').hide(); $('.pagecontent2').hide(); }); // --></script> <div> <div style="float: right; margin-bottom: -20px; margin-left: -20px; width: 265px;"><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/western_blotting/technology_detail/western-blotting-doctor-image.jpg" alt="" width="248" height="165" /></div> <div style="float: left; width: 325px;"> <p>The Western Blot Doctor is a self-help guide developed by Bio-Rad researchers that enables you to identify and troubleshoot western blotting problems. Comprehensive solutions and suggestions are provided to help solve your particular western blotting challenges.</p> </div> <div style="float: left; width: 325px;"> <p>To start troubleshooting your western blotting problems, choose the type of problem you are experiencing.</p> </div> </div> <div class="clear">&nbsp;</div> <div class="contentroundwrap" style="margin-top:6px"> <div class="contentroundtop"> <h3>I am having trouble with:</h3> </div> <div class="contentroundmidwrap"> <div class="contentroundmid"> <table class="link_table" border="0"> <tbody> <tr> <td class="right-padding-15" align="left" valign="top"> <h5><a href="/evportal/destination/solutions?catID=MIW4Q8C4S"><strong>Protein Band Appearance <br /></strong></a></h5> <ul class="rightarrow"> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-appearance-problems#inconsistentcontrol">Inconsistent control protein levels across samples</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-appearance-problems#ghostprotein">Ghost protein bands</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-appearance-problems#swirls">Swirls or missing bands; bands appear diffuse on blot</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-appearance-problems#blurrybands">Blurry bands</a></li> <a href="/en-us/applications-technologies/western-blot-doctor-protein-band-appearance-problems#blurrybands"> </a> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-appearance-problems#blurrybands"></a><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-appearance-problems#smiling">Bands are curved (smiling) not straight</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-appearance-problems#broadband">Broad or misshapen bands</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-appearance-problems#whitebands">White (negative) bands on film using ECL method</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-appearance-problems#nobands">No bands</a></li> </ul> <h5><a href="/evportal/destination/solutions?catID=MIW4O8ESH"><strong>Blot Background <br /></strong></a></h5> <ul class="rightarrow"> <li><a href="/en-us/applications-technologies/western-blot-doctor-blot-background-problems#whitespots">White spots or regions on membrane</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-blot-background-problems#highbackground">High background on blot</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-blot-background-problems#blotchybackground">Blotchy or patchy background</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-blot-background-problems#speckledbackground">Uneven spots on blot or speckled background</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-blot-background-problems#tankblotters">Shadow of tank blotter gel cassette on blot</a></li> </ul> </td> <td align="left" valign="top"> <h5><a href="/evportal/destination/solutions?catID=MIW4S08UU"><strong>Signal Strength</strong></a></h5> <ul class="rightarrow"> <li><a href="/en-us/applications-technologies/western-blot-doctor-signal-strength-problems#uneven">Uneven signal across blot</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-signal-strength-problems#targetsignal">Saturated band signal</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-signal-strength-problems#fainthighmwbands">Faint high-MW bands</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-signal-strength-problems#weaknosignal">Faint bands, weak or no signal</a></li> </ul> <h5><a href="/evportal/destination/solutions?catID=MIW4MCKG4"><strong>Protein Band Size and Pattern</strong></a></h5> <ul class="rightarrow"> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-size-pattern-problems#lowermw">Band(s) at lower MW than expected</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-size-pattern-problems#slightlyhighermw">Band(s) at slightly higher MW than expected, and may be blurry</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-size-pattern-problems#significantlyhighermw">Band(s) at significantly higher MW than expected</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-size-pattern-problems#uneventransfer">Multiple bands at various MWs</a></li> </ul> </td> </tr> </tbody> </table> </div> <div class="contentroundbot">&nbsp;</div> </div> </div> <br /> <h2 class="descHeader">Related Topics:</h2> <p><a href="/evportal/destination/solutions?catID=LUSPPSESH" target="_self">Protein Blotting Methods</a>, <a href="/evportal/destination/solutions?catID=LUSPQXC4S" target="_self">Protein Blotting Equipment (Cells and Power Supplies)</a>, <a href="/evportal/destination/solutions?catID=LUSPSJ8UU" target="_self">Membranes and Blotting Papers</a>, <a href="/evportal/destination/solutions?catID=LUSPTIMNI" target="_self">Transfer Conditions</a>, <a href="/evportal/destination/solutions?catID=LUSPULKSY" target="_self">Protein Blotting Detection and Imaging</a>, <a href="/evportal/destination/solutions?catID=LUSQA88UU" target="_self">Transfer Buffers</a></p> <p>Good western blotting results begin with high-quality protein electrophoresis results. For detailed descriptions of the best practices for optimal protein separation by polyacrylamide gel electrophoresis, visit the <a href="/evportal/destination/solutions?catID=LUSOVO47B">Protein Electrophoresis</a> section.</p> <p><a href="#Relatedcontent">Download individual western blotting protocols</a> in PDF format in the Protocols tab below.</p> <br /><br /> Supporting Documents <ul> <li><a href="/webroot/web/pdf/lsr/literature/Bulletin_6040.pdf" target="_blank">Protein Electrophoresis Guide</a></li> <li><a href="/webroot/web/pdf/lsr/literature/Bulletin_2895.pdf" target="_blank">Protein Blotting Guide</a></li> <li><a href="/webroot/web/pdf/lsr/literature/Bulletin_2895.pdf#page=38" target="_blank">Glossary of Western Blotting Terms</a></li> </ul> Protocols <table id="carttablealigned" class="literature_table" style="height: auto; width: 583px;" border="0" cellspacing="0" cellpadding="0"> <tbody> <tr> <td width="100">6359</td> <td width="350">Avoiding Housekeeping Protein Detection Saturation Protocol, Rev A</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6359.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td>6361</td> <td>Determining the Appropriate Film Exposure Time Protocol, Rev A</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6361.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td width="100">6362</td> <td width="350">Determining the Appropriate Sample Load for Western Blots Protocol, Rev A</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6362.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td>6363</td> <td>Determining the Appropriate Sample Load When Using a Stain-Free V3 Western Workflow&trade; Protocol, Rev A</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6363.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td>6366</td> <td>Validating the Expression Consistency of a Housekeeping Protein Protocol, Rev A</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6366.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td>6376</td> <td>General Protocol for Western Blotting Protocol, Rev A</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6376.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td>6390</td> <td>General V3 Western Workflow Blotting Protocol, Rev A</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6390.pdf" target="_blank"><span>Click to download</span></a></td> </tr> </tbody> </table> <div class="videowrap"> <div class="videoImg"><a title="Total Protein Normalization on Western Blots" href="javascript:openAjaxOverlay('/webroot/web/html/lsr/products/electrophoresis/product_overlay/global/total-protein-normalization.html');"><img style="width: 88px; 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height: 51px; border: 0px none;" src="/webroot/web/images/lsr/products/electrophoresis/product_overlay_icons/global/v3-trans-blot-turbo-transfer-system-faster-western-blotting.jpg" alt="Trans-Blot Turbo System Introduction" /></a></div> <div class="videoDesc"><a title="Trans-Blot Turbo Transfer System: Faster Western Blotting" onclick="javascript:openElementOverlay('tbtWb');" href="javascript:void(0);">Trans-Blot<sup>&reg;</sup> Turbo&trade; Transfer System: Faster Western Blotting</a> <br /></div> <div class="clear">&nbsp;</div> </div> <div id="tbtWb" style="display: none;"> <div class="overlay_contents" style="height: 383px;"> <div class="overlay-head">Trans-Blot<sup>&reg;</sup> Turbo&trade; Transfer System: Faster Western Blotting</div> <iframe src="http://www.youtube.com/embed/laqGU0RpQAo?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" width="620" height="376"></iframe></div> </div> 6517 /templatedata/internet/documentation/data/LSR/Literature/6517_1410803267.xml ChemiDoc Touch Imaging System Brochure, Ver D 6517 /webroot/web/pdf/lsr/literature/Bulletin_6517.pdf Literature PDF Brochures_and_Specifications ChemiDoc Touch Imaging System Brochure No ChemiDoc&trade; Touch Imaging System Brochure, Ver D 6517 6517, western, blotting, image, lab, touch, stain-free, stainfree, smart, tray, technology, chemiluminescent, v3, workflow, total, protein, normalization, tpn, surebeads, tgx, miniprotean, mini-protean, criterion, fastcast, trans-blot, turbo, transblot, tbt, precision, plus, protein, precisionab, antibodies, clarity, max, ECL, substrate, 1708370, 1708381, 1708382, 1709690 6427 /templatedata/internet/documentation/data/LSR/Literature/6427.xml Bio-Rad<sup>&reg;</sup> V3 Western Workflow&trade;, Ver D 6427 H /webroot/web/pdf/lsr/literature/Bulletin_6427.pdf Literature PDF Brochures_and_Specifications /webroot/web/images/general/icons/icon_pdf.gif Bio-Rad V3 Western Workflow No Bio-Rad<sup>&reg;</sup> V3 Western Workflow&trade;, Ver D 6427 6427, bulletin, v3 western workflow, western blot, immunoblot, stain-free, stain free, gel, mini-protean, mini, protean, criterion 6349 /templatedata/internet/documentation/data/LSR/Literature/6349.xml 6350 /templatedata/internet/documentation/data/LSR/Literature/6350.xml Housekeeping Protein Normalization Publications, Rev A 6350 /webroot/web/pdf/lsr/literature/Bulletin_6350.pdf Literature PDF Articles_and_Whitepapers /webroot/web/images/general/icons/icon_pdf.gif Housekeeping Protein Normalization Publications No Housekeeping Protein Normalization Publications, Rev A 6350 Housekeeping Protein Normalization Publications 6351 /templatedata/internet/documentation/data/LSR/Literature/6351.xml Selected Publications List: Total Protein Normalization in Western Blotting Using Stain-Free Technology 6351 /webroot/web/pdf/lsr/literature/Bulletin_6351.pdf Literature PDF Other /webroot/web/images/general/icons/icon_pdf.gif Selected Publications List: Total Protein Normalization in Western Blotting Using Stain-Free Technology No Selected Publications List: Total Protein Normalization in Western Blotting Using Stain-Free Technology 6351 stain free, tgx, stainfree, stain-free, blot, blotting, gel, imaging, gel imaging, blot imaging, western blotting, protein normalization, tpn, housekeeping, loading control, v3 western workflow 6356 /templatedata/internet/documentation/data/LSR/Literature/6356.xml Traditional vs. V3 Western Workflow 6356 /webroot/web/pdf/lsr/literature/Bulletin_6356.pdf Literature PDF Other /webroot/web/images/general/icons/icon_pdf.gif Traditional vs. V3 Western Workflow No Traditional vs. V3 Western Workflow 6356 Traditional, V3, Western, Workflow 6360 /templatedata/internet/documentation/data/LSR/Literature/6360.xml A Method for Greater Reliability in Western Blot Loading Controls: Stain-Free Total Protein Quantitation, Rev B 6360 /webroot/web/pdf/lsr/literature/Bulletin_6360.pdf Literature PDF Application_Notes /webroot/web/images/general/icons/icon_pdf.gif A Method for Greater Reliability in Western Blot Loading Controls: Stain-Free Total Protein Quantitation No A Method for Greater Reliability in Western Blot Loading Controls: Stain-Free Total Protein Quantitation, Rev B 6360 Method for Greater Reliability, Western Blot, Loading Controls, Stain-Free, Total Protein Quantitation RP0058 /templatedata/internet/documentation/data/LSR/Literature/RP0058_1381251352.xml A Defined Methodology for Reliable Quantification of Western Blot Data RP0058 /webroot/web/pdf/lsr/literature/RP0058.pdf Literature PDF Articles_and_Whitepapers /webroot/web/images/general/icons/icon_pdf.gif A Defined Methodology for Reliable Quantification of Western Blot Data No A Defined Methodology for Reliable Quantification of Western Blot Data RP0058 western blotting, western blot, densitometry, protein expression, normalization, quantitation, quantitative, stain-free imaging, stain-free gel, stain-free blot, stain-free blotting, housekeeping protein, housekeeping proteins, controls, densitometry, protein expression Karen Moss Troubleshooting Western Blots western, blot, blotting, doctor, troubleshoot, troubleshooting, self-help, guide 02/27/13 05:34 PM 02/27/23 05:36 PM AE,AI,AL,AM,AR,AT,AU,AZ,BA,BD,BE,BF,BG,BH,BN,BO,BR,BW,CA,CH,CL,CM,CN,CO,CR,CY,CZ,DE,DK,DO,DZ,EC,EE,EG,EH,ER,ES,ET,FI,FM,FO,FR,GA,GE,GF,GH,GP,GR,GT,GU,HK,HN,HR,HT,HU,ID,IE,IL,IN,IS,IT,JM,JO,JP,KE,KH,KR,KW,KZ,LB,LI,LK,LT,LU,LV,MA,MD,MG,MK,ML,MO,MQ,MS,MT,MU,MX,MY,NG,NI,NL,NO,NP,NZ,OM,PA,PE,PF,PG,PH,PK,PL,PR,PS,PT,PW,PY,QA,RO,RS,RU,SA,SB,SE,SG,SI,SK,SN,ST,SV,TG,TH,TN,TO,TR,TT,TW,TZ,UA,UG,UK,US,UY,UZ,VA,VE,VU,XK,YE,ZA en LSR /LSR/Technologies/Western_Blotting N 0 /en-us/applications-technologies/url-title-missing?ID=MIW4JGE8Z /en-us/applications-technologies/url-title-missing?ID=MIW4KT4VY Western Blot Doctor&#153; — Protein Band Size and Pattern Problems /en-us/applications-technologies/applications-technologies/western-blot-doctor-protein-band-size-pattern-problems?ID=MIW4MCKG4 Western Blot Doctor&#153; — Blot Background Problems /en-us/applications-technologies/applications-technologies/western-blot-doctor-blot-background-problems?ID=MIW4O8ESH Western Blot Doctor&#153; — Protein Band Appearance Problems /en-us/applications-technologies/applications-technologies/western-blot-doctor-protein-band-appearance-problems?ID=MIW4Q8C4S Western Blot Doctor&#153; — Signal Strength Problems /en-us/applications-technologies/applications-technologies/western-blot-doctor-signal-strength-problems?ID=MIW4S08UU Introduction to Western Blotting /en-us/applications-technologies/applications-technologies/troubleshooting-western-blots-with-western-blot-doctor?ID=LUSPPAKG4

The Western Blot Doctor is a self-help guide developed by Bio-Rad researchers that enables you to identify and troubleshoot western blotting problems. Comprehensive solutions and suggestions are provided to help solve your particular western blotting challenges.

To start troubleshooting your western blotting problems, choose the type of problem you are experiencing.

 

Related Topics:

Protein Blotting Methods, Protein Blotting Equipment (Cells and Power Supplies), Membranes and Blotting Papers, Transfer Conditions, Protein Blotting Detection and Imaging, Transfer Buffers

Good western blotting results begin with high-quality protein electrophoresis results. For detailed descriptions of the best practices for optimal protein separation by polyacrylamide gel electrophoresis, visit the Protein Electrophoresis section.

Download individual western blotting protocols in PDF format in the Protocols tab below.



 
 
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Literature
Number Description Download
6517 ChemiDoc Touch Imaging System Brochure, Ver D Click to download
6427 Bio-Rad<sup>&reg;</sup> V3 Western Workflow&trade;, Ver D Click to download
6350 Housekeeping Protein Normalization Publications, Rev A Click to download
6351 Selected Publications List: Total Protein Normalization in Western Blotting Using Stain-Free Technology Click to download
6356 Traditional vs. V3 Western Workflow Click to download
6360 A Method for Greater Reliability in Western Blot Loading Controls: Stain-Free Total Protein Quantitation, Rev B Click to download
RP0058 A Defined Methodology for Reliable Quantification of Western Blot Data Click to download
6359 Avoiding Housekeeping Protein Detection Saturation Protocol, Rev A Click to download
6361 Determining the Appropriate Film Exposure Time Protocol, Rev A Click to download
6362 Determining the Appropriate Sample Load for Western Blots Protocol, Rev A Click to download
6363 Determining the Appropriate Sample Load When Using a Stain-Free V3 Western Workflow™ Protocol, Rev A Click to download
6366 Validating the Expression Consistency of a Housekeeping Protein Protocol, Rev A Click to download
6376 General Protocol for Western Blotting Protocol, Rev A Click to download
6390 General V3 Western Workflow Blotting Protocol, Rev A Click to download
 
 
MIW4HR15 [referer] = [https://www.google.com/] [x-forwarded-proto] = [https] [cookie] = [JSESSIONID=2E9DA1757049B638B8EA5E0D081D422D.lsds-e1c; JSESSIONID_STERLING=F3908D28B8A658A01F77979C2D895869.ecommerce1; mindsparktb_207743773="true"; mindsparktb_226333724="true"; PreviouslyViewedCookie=2e811946-ca88-44bd-8032-d74ca88089a9; SampleCookie=SampleCookie; _4c_=jZPNbts6EIVfpeCiq9AW%2F35kICiKdJNFu8kF7tKgyKEsRBZdSq5qBH73DBWnKZK0qAAKFPXpzJlD6oHMOxjIhikhZWW51ZUWV%2BQeTiPZPBB%2FKPcf5XbMPdmQ3TQdNuv1PM%2Brpks0u7Dyab%2BGgR7HtXcTtCmf1jOME%2BSBNn2apm5oP91%2Bua4FlzEIT7WLkkqlGLWCAbWh0dGJKKTiH%2F%2F%2FbzVmv7tmZbb32y5cu5kevgf67TO9qG6L6raFAbLrt23qQ4Hvh3FXcOY9xAbF6ygdlY3VFIegYIThYI2PXJEr4lMAbIfVK7Wq8Pl2KNKodwcODeArXLxJx2HKp5snFheanOYRMj7cuehy94FVK4brEWMiCgvw2EAtPVcWlAnSRxOENJXk4ARyCUMlX53HaYYIOS9aJdPxEmqbUtvDkilCYzeVyr8ljYtodF90cHrAnSHFQJ%2B86wuKe1m0D707YRqLK%2BHrGFXUJeCGy9o4X0vWBBkh8LpUgXHs0vCEm6oxShlvpG%2B4QJBzG7RULlrHwJHzFfl5OS611FZoXqOlCc%2BG1bIqFxK5aC3nhngdo2GNdpwHUQfHmHGVrni0HLBOSWXRk5xZya3gSqPAAfWW79lLOc6xHI5LOYbn9VKuxPCaXsxxw9%2Bae9rEv3wj32nIP%2Fcz5SO88WzqwkwXJrp%2BhFcIdoyIH%2FxzY3%2BG9i%2Fh%2FcPmvSPQ5vhs5ddvja%2Fwt34Fi%2FP5%2FAg%3D; _mkto_trk=id:272-THL-329&token:_mch-bio-rad.com-1534408282342-60362; WT_FPC=id=910aee1c-2e9c-4836-85ca-ef8cbf2059f1:lv=1534393882675:ss=1534393882675] [accept-language] = [es-xl] [x-forwarded-port] = [443] [x-forwarded-for] = [24.50.227.163, 10.232.18.193] [accept] = [text/html,application/xhtml+xml,application/xml;q=0.9,*/*;q=0.8] [seourl] = [/en-us/applications-technologies/troubleshooting-western-blots-with-western-blot-doctor] [x-amzn-trace-id] = [Root=1-5b753695-7b3bd97cc29bad36eb122734] [x-forwarded-server] = [lsds-prod-s.br.aws-livesite.io] [x-forwarded-host] = [www.bio-rad.com] [x-query-string] = [ID=MIW4HR15&WT.srch=1&WT.mc_id=&WT.knsh_id=] [host] = [10.232.1.21:1776] [x-request-uri] = [/en-us/applications-technologies/troubleshooting-western-blots-with-western-blot-doctor] [connection] = [Keep-Alive] [accept-encoding] = [gzip, deflate] [user-agent] = [Mozilla/5.0 (Macintosh; Intel Mac OS X 10_10_5) AppleWebKit/603.3.8 (KHTML, like Gecko) Version/10.1.2 Safari/603.3.8] 1 AppTech/AppTechDetails pageStyleKey internet/solutions_sub applications-technologies/troubleshooting-western-blots-with-western-blot-doctor LSR MIW4HR15 Western Blot Doctor Troubleshooting Western Blots with the Western Blot Doctor&#153; /webroot/web/html/lsr/solutions/technologies/western_blotting <script type="text/javascript"><!-- $(document).ready(function(){ $('.analysismethods').hide(); $('.pagecontent2').hide(); }); // --></script> <div> <div style="float: right; margin-bottom: -20px; margin-left: -20px; width: 265px;"><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/western_blotting/technology_detail/western-blotting-doctor-image.jpg" alt="" width="248" height="165" /></div> <div style="float: left; width: 325px;"> <p>The Western Blot Doctor is a self-help guide developed by Bio-Rad researchers that enables you to identify and troubleshoot western blotting problems. Comprehensive solutions and suggestions are provided to help solve your particular western blotting challenges.</p> </div> <div style="float: left; width: 325px;"> <p>To start troubleshooting your western blotting problems, choose the type of problem you are experiencing.</p> </div> </div> <div class="clear">&nbsp;</div> <div class="contentroundwrap" style="margin-top:6px"> <div class="contentroundtop"> <h3>I am having trouble with:</h3> </div> <div class="contentroundmidwrap"> <div class="contentroundmid"> <table class="link_table" border="0"> <tbody> <tr> <td class="right-padding-15" align="left" valign="top"> <h5><a href="/evportal/destination/solutions?catID=MIW4Q8C4S"><strong>Protein Band Appearance <br /></strong></a></h5> <ul class="rightarrow"> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-appearance-problems#inconsistentcontrol">Inconsistent control protein levels across samples</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-appearance-problems#ghostprotein">Ghost protein bands</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-appearance-problems#swirls">Swirls or missing bands; bands appear diffuse on blot</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-appearance-problems#blurrybands">Blurry bands</a></li> <a href="/en-us/applications-technologies/western-blot-doctor-protein-band-appearance-problems#blurrybands"> </a> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-appearance-problems#blurrybands"></a><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-appearance-problems#smiling">Bands are curved (smiling) not straight</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-appearance-problems#broadband">Broad or misshapen bands</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-appearance-problems#whitebands">White (negative) bands on film using ECL method</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-appearance-problems#nobands">No bands</a></li> </ul> <h5><a href="/evportal/destination/solutions?catID=MIW4O8ESH"><strong>Blot Background <br /></strong></a></h5> <ul class="rightarrow"> <li><a href="/en-us/applications-technologies/western-blot-doctor-blot-background-problems#whitespots">White spots or regions on membrane</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-blot-background-problems#highbackground">High background on blot</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-blot-background-problems#blotchybackground">Blotchy or patchy background</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-blot-background-problems#speckledbackground">Uneven spots on blot or speckled background</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-blot-background-problems#tankblotters">Shadow of tank blotter gel cassette on blot</a></li> </ul> </td> <td align="left" valign="top"> <h5><a href="/evportal/destination/solutions?catID=MIW4S08UU"><strong>Signal Strength</strong></a></h5> <ul class="rightarrow"> <li><a href="/en-us/applications-technologies/western-blot-doctor-signal-strength-problems#uneven">Uneven signal across blot</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-signal-strength-problems#targetsignal">Saturated band signal</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-signal-strength-problems#fainthighmwbands">Faint high-MW bands</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-signal-strength-problems#weaknosignal">Faint bands, weak or no signal</a></li> </ul> <h5><a href="/evportal/destination/solutions?catID=MIW4MCKG4"><strong>Protein Band Size and Pattern</strong></a></h5> <ul class="rightarrow"> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-size-pattern-problems#lowermw">Band(s) at lower MW than expected</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-size-pattern-problems#slightlyhighermw">Band(s) at slightly higher MW than expected, and may be blurry</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-size-pattern-problems#significantlyhighermw">Band(s) at significantly higher MW than expected</a></li> <li><a href="/en-us/applications-technologies/western-blot-doctor-protein-band-size-pattern-problems#uneventransfer">Multiple bands at various MWs</a></li> </ul> </td> </tr> </tbody> </table> </div> <div class="contentroundbot">&nbsp;</div> </div> </div> <br /> <h2 class="descHeader">Related Topics:</h2> <p><a href="/evportal/destination/solutions?catID=LUSPPSESH" target="_self">Protein Blotting Methods</a>, <a href="/evportal/destination/solutions?catID=LUSPQXC4S" target="_self">Protein Blotting Equipment (Cells and Power Supplies)</a>, <a href="/evportal/destination/solutions?catID=LUSPSJ8UU" target="_self">Membranes and Blotting Papers</a>, <a href="/evportal/destination/solutions?catID=LUSPTIMNI" target="_self">Transfer Conditions</a>, <a href="/evportal/destination/solutions?catID=LUSPULKSY" target="_self">Protein Blotting Detection and Imaging</a>, <a href="/evportal/destination/solutions?catID=LUSQA88UU" target="_self">Transfer Buffers</a></p> <p>Good western blotting results begin with high-quality protein electrophoresis results. For detailed descriptions of the best practices for optimal protein separation by polyacrylamide gel electrophoresis, visit the <a href="/evportal/destination/solutions?catID=LUSOVO47B">Protein Electrophoresis</a> section.</p> <p><a href="#Relatedcontent">Download individual western blotting protocols</a> in PDF format in the Protocols tab below.</p> <br /><br /> Supporting Documents <ul> <li><a href="/webroot/web/pdf/lsr/literature/Bulletin_6040.pdf" target="_blank">Protein Electrophoresis Guide</a></li> <li><a href="/webroot/web/pdf/lsr/literature/Bulletin_2895.pdf" target="_blank">Protein Blotting Guide</a></li> <li><a href="/webroot/web/pdf/lsr/literature/Bulletin_2895.pdf#page=38" target="_blank">Glossary of Western Blotting Terms</a></li> </ul> Protocols <table id="carttablealigned" class="literature_table" style="height: auto; width: 583px;" border="0" cellspacing="0" cellpadding="0"> <tbody> <tr> <td width="100">6359</td> <td width="350">Avoiding Housekeeping Protein Detection Saturation Protocol, Rev A</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6359.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td>6361</td> <td>Determining the Appropriate Film Exposure Time Protocol, Rev A</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6361.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td width="100">6362</td> <td width="350">Determining the Appropriate Sample Load for Western Blots Protocol, Rev A</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6362.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td>6363</td> <td>Determining the Appropriate Sample Load When Using a Stain-Free V3 Western Workflow&trade; Protocol, Rev A</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6363.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td>6366</td> <td>Validating the Expression Consistency of a Housekeeping Protein Protocol, Rev A</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6366.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td>6376</td> <td>General Protocol for Western Blotting Protocol, Rev A</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6376.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td>6390</td> <td>General V3 Western Workflow Blotting Protocol, Rev A</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6390.pdf" target="_blank"><span>Click to download</span></a></td> </tr> </tbody> </table> <div class="videowrap"> <div class="videoImg"><a title="Total Protein Normalization on Western Blots" href="javascript:openAjaxOverlay('/webroot/web/html/lsr/products/electrophoresis/product_overlay/global/total-protein-normalization.html');"><img style="width: 88px; height: 51px; border: 0px none;" src="/webroot/web/images/lsr/products/electrophoresis/product_overlay_icons/global/tpn-on-western-blots-thumb.jpg" alt="Total Protein Normalization on Western Blots" /></a></div> <div class="videoDesc"><a title="Total Protein Normalization on Western Blots" href="javascript:openAjaxOverlay('/webroot/web/html/lsr/products/electrophoresis/product_overlay/global/total-protein-normalization.html');">Using Image Lab&trade; Software for Total Protein Normalization on Western Blots</a> <br /></div> <div class="clear">&nbsp;</div> </div> <div class="videowrap"> <div class="videoImg"><a title="ChemiDoc Touch Testimonial" href="javascript:openAjaxOverlay('/webroot/web/html/lsr/products/imaging_bioinformatics/product_overlay/global/ov-chemidoc-testimonial.html');"><img style="width: 88px; height: 51px; border: 0px none;" src="/webroot/web/images/lsr/products/imaging_bioinformatics/product_overlay_icons/global/ov-chemidoc-testimonial-thumb.jpg" alt="ChemiDoc Touch Testimonial" /></a></div> <div class="videoDesc"><a title="ChemiDoc Touch Testimonial" href="javascript:openAjaxOverlay('/webroot/web/html/lsr/products/imaging_bioinformatics/product_overlay/global/ov-chemidoc-testimonial.html');">ChemiDoc&trade; Touch Testimonial</a> <br /></div> <div class="clear">&nbsp;</div> </div> <div class="videowrap"> <div class="videoImg"><a title="Seeing is Believing" href="javascript:openAjaxOverlay('/webroot/web/html/lsr/products/electrophoresis/product_overlay/global/breast-cancer-pathways-ov.html');"><img style="width: 88px; height: 51px; border: 0px none;" src="/webroot/web/images/lsr/products/electrophoresis/product_overlay_icons/global/breast-cancer-pathways-ov-th.jpg" alt="Seeing is Believing" /></a></div> <div class="videoDesc"><a title="Seeing is Believing" href="javascript:openAjaxOverlay('/webroot/web/html/lsr/products/electrophoresis/product_overlay/global/breast-cancer-pathways-ov.html');">"Seeing is Believing" &mdash; V3 Western Workflow&trade; with Stain-Free Gel Technology Tutorial </a> <br /></div> <div class="clear">&nbsp;</div> </div> <div class="videowrap"> <div class="videoImg"><a title="Can We Trust Western Blots?" href="javascript:openAjaxOverlay('/webroot/web/html/lsr/products/electrophoresis/product_overlay/global/ov-trust-western-blots.html');"><img style="width: 88px; height: 51px; border: 0px none;" src="/webroot/web/images/lsr/products/electrophoresis/product_overlay_icons/global/ov-trust-western-blots-thumb.jpg" alt="Can We Trust Western Blots?" /></a></div> <div class="videoDesc"><a title="Can We Trust Western Blots?" href="javascript:openAjaxOverlay('/webroot/web/html/lsr/products/electrophoresis/product_overlay/global/ov-trust-western-blots.html');">"Can We Trust Western Blots?" Webinar</a> <br /></div> <div class="clear">&nbsp;</div> </div> <div class="videowrap"> <div class="videoImg"><a title="Single Cell Western Blot and Stain-Free Total PRotein Loading Control" href="javascript:openAjaxOverlay('/webroot/web/html/lsr/products/electrophoresis/product_overlay/global/ov-total-protein-normalization-westerns.html');"><img style="width: 88px; height: 51px; border: 0px none;" src="/webroot/web/images/lsr/products/electrophoresis/product_overlay_icons/global/ov-total-protein-normalization-westerns-thumb.jpg" alt="Single Cell Western Blot and Stain-Free Total PRotein Loading Control" /></a></div> <div class="videoDesc"><a title="Single Cell Western Blot and Stain-Free Total PRotein Loading Control" href="javascript:openAjaxOverlay('/webroot/web/html/lsr/products/electrophoresis/product_overlay/global/ov-total-protein-normalization-westerns.html');">"Single Cell Western Blot and Stain-Free Total Protein Loading Control" Webinar</a> <br /></div> <div class="clear">&nbsp;</div> </div> <div class="videowrap"> <div class="videoImg"><a title="Trans-Blot Turbo System Introduction" onclick="javascript:openElementOverlay('tbtIntro');" href="javascript:void(0);"><img style="width: 88px; height: 51px; border: 0px none;" src="/webroot/web/images/lsr/products/electrophoresis/product_overlay_icons/global/tbt_systour.jpg" alt="Trans-Blot Turbo System Introduction" /></a></div> <div class="videoDesc"><a title="Trans-Blot Turbo System Introduction" onclick="javascript:openElementOverlay('tbtIntro');" href="javascript:void(0);">Trans-Blot<sup>&reg;</sup> Turbo&trade; Transfer System Introduction</a> <br /></div> <div class="clear">&nbsp;</div> </div> <div id="tbtIntro" style="display: none;"> <div class="overlay_contents" style="height: 383px;"> <div class="overlay-head">Trans-Blot<sup>&reg;</sup> Turbo&trade; Transfer System Introduction</div> <iframe src="http://www.youtube.com/embed/lV10IYOz29A?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" width="620" height="376"></iframe></div> </div> <div class="videowrap"> <div class="videoImg"><a title="Get to Know the Trans-Blot Turbo Transfer System" onclick="javascript:openAjaxOverlay('/webroot/web/html/lsr/products/electrophoresis/product_overlay/global/tbt_videos.html' );" href="javascript:void(0);"><img style="border: medium none;" src="/webroot/web/images/lsr/products/electrophoresis/product_overlay_icons/global/TBT_vid01_instructional.jpg" alt="Get to Know the Trans-Blot Turbo Transfer System" /></a></div> <div class="videoDesc"><a title="Get to Know the Trans-Blot Turbo Transfer System" onclick="javascript:openAjaxOverlay('/webroot/web/html/lsr/products/electrophoresis/product_overlay/global/tbt_videos.html' );" href="javascript:void(0);">Get to Know the Trans-Blot<sup>&reg;</sup> Turbo&trade; Transfer System</a><br /></div> <div class="clear">&nbsp;</div> </div> <div class="videowrap"> <div class="videoImg"><a title="Trans-Blot Turbo System vs. Traditional Blotting Techniques" onclick="javascript:openAjaxOverlay('/webroot/web/html/lsr/products/electrophoresis/product_overlay/global/tbt_video_02.html' );" href="javascript:void(0);"><img style="border: medium none;" src="/webroot/web/images/lsr/products/electrophoresis/product_overlay_icons/global/TBT_vid02_vsTrad.jpg" alt="Trans-Blot Turbo System vs. Traditional Blotting Techniques" /></a></div> <div class="videoDesc"><a title="Trans-Blot Turbo System vs. Traditional Blotting Techniques" onclick="javascript:openAjaxOverlay('/webroot/web/html/lsr/products/electrophoresis/product_overlay/global/tbt_video_02.html' );" href="javascript:void(0);">Trans-Blot<sup>&reg;</sup> Turbo&trade; System vs. Traditional Blotting Techniques</a><br /></div> <div class="clear">&nbsp;</div> </div> <div class="videowrap"> <div class="videoImg"><a title="Trans-Blot Turbo Transfer System: Faster Western Blotting" onclick="javascript:openElementOverlay('tbtWb');" href="javascript:void(0);"><img style="width: 88px; height: 51px; border: 0px none;" src="/webroot/web/images/lsr/products/electrophoresis/product_overlay_icons/global/v3-trans-blot-turbo-transfer-system-faster-western-blotting.jpg" alt="Trans-Blot Turbo System Introduction" /></a></div> <div class="videoDesc"><a title="Trans-Blot Turbo Transfer System: Faster Western Blotting" onclick="javascript:openElementOverlay('tbtWb');" href="javascript:void(0);">Trans-Blot<sup>&reg;</sup> Turbo&trade; Transfer System: Faster Western Blotting</a> <br /></div> <div class="clear">&nbsp;</div> </div> <div id="tbtWb" style="display: none;"> <div class="overlay_contents" style="height: 383px;"> <div class="overlay-head">Trans-Blot<sup>&reg;</sup> Turbo&trade; Transfer System: Faster Western Blotting</div> <iframe src="http://www.youtube.com/embed/laqGU0RpQAo?version=3&amp;rel=0&amp;showinfo=0&amp;theme=light&amp;modestbranding=1&amp;fs=1;wmode=transparent" width="620" height="376"></iframe></div> </div> 6517 /templatedata/internet/documentation/data/LSR/Literature/6517_1410803267.xml 6427 /templatedata/internet/documentation/data/LSR/Literature/6427.xml 6349 /templatedata/internet/documentation/data/LSR/Literature/6349.xml 6350 /templatedata/internet/documentation/data/LSR/Literature/6350.xml 6351 /templatedata/internet/documentation/data/LSR/Literature/6351.xml 6356 /templatedata/internet/documentation/data/LSR/Literature/6356.xml 6360 /templatedata/internet/documentation/data/LSR/Literature/6360.xml RP0058 /templatedata/internet/documentation/data/LSR/Literature/RP0058_1381251352.xml Karen Moss Troubleshooting Western Blots western, blot, blotting, doctor, troubleshoot, troubleshooting, self-help, guide 02/27/13 05:34 PM 02/27/23 05:36 PM AE,AI,AL,AM,AR,AT,AU,AZ,BA,BD,BE,BF,BG,BH,BN,BO,BR,BW,CA,CH,CL,CM,CN,CO,CR,CY,CZ,DE,DK,DO,DZ,EC,EE,EG,EH,ER,ES,ET,FI,FM,FO,FR,GA,GE,GF,GH,GP,GR,GT,GU,HK,HN,HR,HT,HU,ID,IE,IL,IN,IS,IT,JM,JO,JP,KE,KH,KR,KW,KZ,LB,LI,LK,LT,LU,LV,MA,MD,MG,MK,ML,MO,MQ,MS,MT,MU,MX,MY,NG,NI,NL,NO,NP,NZ,OM,PA,PE,PF,PG,PH,PK,PL,PR,PS,PT,PW,PY,QA,RO,RS,RU,SA,SB,SE,SG,SI,SK,SN,ST,SV,TG,TH,TN,TO,TR,TT,TW,TZ,UA,UG,UK,US,UY,UZ,VA,VE,VU,XK,YE,ZA en LSR /LSR/Technologies/Western_Blotting N 0
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