Your Comprehensive Resource for Fluorescent Western Blotting
Achieve Better Data Every Time with Bio-Rad’s Validated Fluorescent Blotting Solutions
ChemiDoc™ Pro Imaging System
Precision You Can Trust, Data You Can Publish
See more from every sample with fluorescent western blotting using the ChemiDoc Pro Imaging System. Confidently image chemiluminescent and fluorescent blots, Stain-Free gels and blots, and all common nucleic acid and protein gel stains. The ChemiDoc Pro System utilizes advanced CMOS digital imaging to capture gel and western blot images with high sensitivity, helping reveal both faint and intense bands. Maximize sample value with 4-plex imaging and custom applications. Connect the system to the BR.io Cloud Platform for easy image upload, access, automated image analysis, and storage.
The ChemiDoc Pro Imaging System mentioned on this webpage is for Research Use Only. Not for use in diagnostic procedures.
Choose the Dyes You Want, Get the Flexibility You Need
UV
- Ethidium bromide
- Oriole
- Stain-Free
- SYBR® Green
- SYBR® Safe
- SYBR® Gold
- Flamingo
Blue
- DyLight 488
- Alexa Fluor 488
- Cy2
- Fluorescein (FITC)
- SYBR® Green
- SYBR® Safe
- SYBR® Gold
- StarBright™
Blue 520 - StarBright
Blue 700
Green
- DyLight 550
- Alexa Fluor 546
- Cy3
- hFAB Rhodamine
- StarBright
Yellow 605
Red
- DyLight 650
- Alexa Fluor 647
- Cy5
- SYPRO Ruby
Far Red
- DyLight 680
- Alexa Fluor 680
- Cy5.5
- IRDye 680
- StarBright
Red 815
Near Infrared
- DyLight 800
- Alexa Fluor 790
- Cy7
- IRDye 800
Bolded fluorophores are available from Bio-Rad.
Discover 10 Tips for Western Blot Detection of Phosphorylation Events
Light Up Your Lab, Simplify Your Selection
Get a free excitation and emission reference poster to make it easier to select the right fluorophore for your next experiment.
Benefits of ChemiDoc Imaging Systems
With the ChemiDoc Pro System, one instrument supports your gel documentation, chemiluminescence, and multiplex fluorescence needs.
Stain-Free
Total Protein Normalization
Learn how to properly design a quantitative western blot experiment and faithfully compare protein expression levels without concern for variable or overexpressed housekeeping proteins.
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.
Multiplex Fluorescence
Up to Four Channels
Conserve sample, avoid errors commonly associated with stripping, reprobing, and cutting blots, and answer more complex biological questions with multiplex fluorescence western blotting.
Better Data
See up to four signals at once or look at a subset of signals. Plus, you can normalize beyond a single protein with Stain-Free total protein normalization.
Explore blot combinations by making a selection on the right.
Image Lab Touch Software
Image Lab Touch Software works with the ChemiDoc Pro System and provides easy and intuitive onboard acquisition software. Built for labs with multiple users, Image Lab Touch Software teaches as you tap and explore, selects optimal light source, and permits both manual entry and automated calculation of exposure times.
Stain-Free Western Blotting
Faster Results. Better Data.
Stain-free western blotting allows you to quickly check electrophoresis and blot transfer quality and obtain truly quantitative western blotting results, updating traditional blotting techniques with innovative tools.
Master Stain-Free Workflows & Total Protein Normalization
NEW! Introducing the Stain-Free Western Blotting Resource Collection, a four-part literature series created to help researchers effectively learn and apply Stain-Free western blotting techniques. It provides comprehensive guidance on mastering both Stain-Free workflows and total protein normalization.
Antibodies for Fluorescent Western Blotting
-
Image
New New Phospho-Specific PrecisionAb™ Antibodies
- Highly specific antibodies to detect phosphorylation events
- Validated by comparing phosphorylation levels in both treated versus untreated cell lysates and by dephosphorylating western blot membranes
- Showcased against total protein antibodies
-
Image
PrecisionAb Antibodies
- Highly specific and sensitive
- Validated for western blotting applications
- Each antibody is screened against up to 12 whole-cell lysates expressing endogenous protein levels
-
Image
hFAB Rhodamine Housekeeping Protein Fluorescent Primary Antibodies
- One-step, directly conjugated fluorescence detection solution for commonly used housekeeping proteins (actin, tubulin, and GAPDH)
- Eliminates cross-reactivity with primary and secondary antibodies from other species
- HuCAL technology allows you to choose any appropriate host species for your other antibodies
Secondary Antibodies
-
Image
StarBright Blue 700 Fluorescent Secondary Antibodies
- Best choice for maximum sensitivity
- Novel polymer-based chemistry is equivalent to femto ECL
- Superior signal-to-noise ratio
-
Image
StarBright Blue 520 Fluorescent Secondary Antibodies
- Novel polymer-based chemistry is equivalent to femto ECL
- Ideal for multiplexing
-
Image
DyLight Fluorescent Secondary Antibodies
- Traditional fluorescent solution
- Available in many colors and host species
- Ideal for routine fluorescent detection
Resources
Western Workflow
- The Complete Guide to Stain-Free Western Blotting
- Stain-Free Western Blotting Brochure
- Stain-Free Western Blotting Publication List
- Western Blotting Protocol Library
- Stain-Free Western Blotting Protocol
- Stain-Free Approach to Western Blotting
Imaging Systems and Antibodies
Imaging Systems:
- New Standard of Confidence From Blot to Results: The ChemiDoc™ Pro Imaging System Brochure
Antibodies:
- StarBright Secondary Antibody Brochure
- StarBright Secondary Antibody Protocol
Western Blot University
Build your western blotting expertise with courses designed for every skill level.
Introducing the Western Blot Learning Center
Improve your western blotting skills with expert guidance.
-
Image
Free 26-Lane Electrophoresis and Western Blotting Layout Post-It Pad
Plan, track, and never miss another step.
Western Blot Selector Tool
Find the right products using the free Western Blot Selector Tool
FAQs
Chemiluminescence uses enzyme-substrate reactions to produce light for single-target detection, requiring stripping and reprobing for multiple targets. Multiplex fluorescence detection uses fluorophore-conjugated antibodies at distinct wavelengths, enabling simultaneous detection of multiple proteins on the same blot.
Select fluorophores based on your imaging system's excitation and emission capabilities, the number of targets you need to detect, and spectral separation to minimize crosstalk. For multiplex experiments, choose fluorophores from differing wavelengths to enable simultaneous detection of multiple targets with clear signal separation.
Fluorescent detection works with both PVDF and nitrocellulose membranes. However, low-fluorescence PVDF membranes are recommended for optimal signal-to-noise ratio. Avoid membranes with high autofluorescence, as they can increase background signal and interfere with fluorescence detection.
Stain-Free technology allows total protein visualization without additional staining steps, enabling real-time assessment of transfer efficiency, accurate normalization to total protein instead of housekeeping proteins, and quality control at every stage of the workflow, from gel to membrane.
High background is commonly caused by insufficient blocking, excessive antibody concentrations, or membrane autofluorescence. Use a blocking reagent compatible with your detection system, optimize primary and secondary antibody dilutions, and ensure thorough washing between incubation steps.
Benefits of Fluorescent Western Blotting
Learn more about fluorescent western blotting through our video.
Fluorescent western blotting enables the simultaneous detection of multiple proteins on the same blot, helping conserve samples, reduce workflow steps, and provide greater flexibility for protein analysis.
- Eliminate stripping and reprobing to reduce workflow steps and preserve samples
- Differentiate closely sized or modified proteins using independent detection wavelengths
- Customize experiments with a range of fluorophores for flexible assay design
Documents
Western Workflow
| Number | Description |
|---|---|
| PDF 4052 | ChemiDoc Pro Imaging System Brochure |
| PDF 6427 | Stain-Free Western Blotting Brochure |
| Library | Western Blotting Protocol Library |
| PDF 6390 | Stain-Free Western Blotting Protocol |
| PDF RP0051 | Stain-Free Approach to Western Blotting |
Imaging Systems
| Number | Description |
|---|---|
| PDF 4052 | ChemiDoc Pro Imaging System Brochure |
Antibodies
| Number | Description |
|---|---|
| PDF 6911 | StarBright Secondary Antibody Brochure |
| PDF 10000078050 | StarBright Secondary Antibody Protocol |
