Bio-Rad Stain-Free Western Blotting Workflow
Faster Results, Better Data
Compared to conventional methods, this improved workflow incorporating Stain-Free gel technology saves time and increases
the accuracy and reliability of your western blot results.

Sample Preparation

Data

BenefitOne-stop shop for consumables and reagentsthat span the complete electrophoresis andwestern blotting workflow

1—Electrophoresis (and Imaging)

2

■

Data

Benefit
Visualize separation for all lanes in 1 min

TGX Stain-Free Gel, Criterion Cell, and ChemiDoc™ Goor ChemiDoc MP Imaging System■+■

Stain-Free image of pre-transfer gel

Transfer (and Imaging)

3

Efficient and uniform protein transfer in 3 min■+Stain-Free image of post-transfer gel

Quickly check transfer efficiency■

Stain-Free image of blot

■
■

Data

Easy multiplexing
Use western blot–validated, highly specific PrecisionAb™ Primary
Antibodies and StarBright Blue Fluorescent Secondary Antibodies
to detect multiple targets on the same blot■

No need to strip and reprobe or to cut the blot■

Smart Tray Technology* automatically recognizes your application

Target protein
detection withfluorescence

DataBenefit
More accurate protein normalization

ChemiDoc Go or ChemiDoc MP Imaging System with
Image Lab Software■

Stain-Free blot (from step 4)
for normalization* U.S. patent 8,913,127.

EveryBlot Blocking Buffer provides 5 min blocking and maximum
sensitivity

Compatible with chemiluminescence andfluorescence detection

Target protein
detection withchemiluminescence

6

Verify transfer quality for all lanes in 2 minBenefit

ChemiDoc Go or ChemiDoc MP Imaging System

Image Analysis

Convenience: choose from ready-to-use transfer packs(nitrocellulose or polyvinylidene difluoride [PVDF]) or ready-toassemble kits (nitrocellulose, PVDF, or low fluorescence [LF] PVDF)

Benefit■5

T hroughput: transfer 4 mini or 2 midi gels at once

Data

EveryBlot Blocking Buffer, Clarity™ and Clarity Max
Western ECL Substrates, and StarBright™ Blue
Fluorescent Secondary Antibodies

Image Acquisition

Speed with flexibility: TGX Stain-Free Gel chemistry available inprecast and handcast formats

Benefit■4

 oomassie-like performance with no background variability
Cand no staining/destaining

Data

Trans-Blot™ Turbo Transfer System, and ChemiDoc Goor ChemiDoc MP Imaging System

Immunodetection

 onvenient premixed electrophoresis gel–forming reagents andCbuffers are the perfect solutions for electrophoresis and blottingneeds■Stain-Free technology eliminates inconsistencies of normalizing toHKPs by using total protein normalization (TPN) for more accurate
protein quantitation and western blotting results
Maintain electronic records for U.S. FDA 21 CFR Part 11
compliance with Image Lab Software, Security Edition

FAQs
How does Stain-Free technology work?
The embedded compound in Stain-Free gels is activated by
UV light, and emits a fluorescent signal with sensitivity equivalent
to that of Coomassie staining. The Stain-Free signal transfers
to the proteins during blotting, which enables the assessmentof transfer efficiency and provides a total protein signal for use
in normalization.
How do total protein stains compare to normalizing with ahousekeeping protein (HKP)?
Advantages of total protein stains vs. HKP normalization include:
1. H
 KP expression may change depending on experimental
conditions and sample type. Total protein is more stable andless prone to change with experimental conditions.
2. T otal protein normalization using Stain-Free technologyeliminates the need to strip and reprobe for HKPs. Strippingand reprobing for HKPs is a very time-consuming process,adding hours to detection, whereas Stain-Free total protein
detection can be done in minutes.
3. Compared to HKPs, total protein stains exhibit superior
linearity and reproducibility.

How does Stain-Free fluorescence compare to other total
protein stains for blots?
Stain-Free technology provides higher detection sensitivity anda wider linear dynamic range than other available total protein
stains. Stain-Free technology eliminates staining and destaining
manipulation, ensuring proteins remain intact from gel to membrane.
Other total protein stains, including Ponceau S and SYPRO Ruby,
can introduce errors in the staining/destaining procedure.
What resources are available?
Tips on normalization with Stain-Free technology are available
at bio-rad.com.
Bulletin 3429
The Complete Guide to Stain-Free Western Blotting
Bulletin 6434
Western Blot Normalization Using Image Lab Software
Bulletin 6390
General Stain-Free Western Blotting Protocol
Bulletin 6360	A Method for Greater Reliability in Western Blot Loading
Controls: Stain-Free Total Protein Quantitation
Bulletin 6351Selected Publications List: Total Protein Normalization in		
Western Blotting Using Stain-Free Technology

Tips for good quantitative western blotting are available
at bio-rad.com.
Bulletin 2895

Protein Blotting Guide

Ordering Information
Catalog #Description

Protein Standards
1610373	Precision Plus Protein All Blue Prestained Protein Standards
1610374
Precision Plus Protein Dual Color Standards, 500 µl
1610363	Precision Plus Protein Unstained Protein Standards, Strep-tagged
recombinant, 1 ml
1610377
Precision Plus Protein Dual Xtra Prestained Protein Standards
Buffers1610732	
10x Tris/Glycine/SDS Buffer
1610737
2x Laemmli Sample Buffer
1610747	
4x Laemmli Sample Buffer
1706435
10x Tris Buffered Saline (TBS)Electrophoresis Cells
1656001	Criterion Cell, includes electrophoresis buffer tank, lid with power cables,
3 sample loading guides
1658004	
Mini-PROTEAN™ Tetra Vertical Electrophoresis Cell for Mini Precast
Gels, 4-gel vertical electrophoresis system, includes electrode assembly,
companion running module, tank, lid with power cables, mini cell buffer dam
Blotting Systems
17001917 Trans-Blot Turbo Transfer Starter System, mini PVDF
17001919 Trans-Blot Turbo Transfer Starter System, midi PVDF
17001918 Trans-Blot Turbo Transfer Starter System, mini nitrocellulose
17001915 Trans-Blot Turbo Transfer Starter System, midi nitrocellulose
1704156	Trans-Blot Turbo Mini 0.2 µm PVDF Transfer Packs, pkg of 101704157	Trans-Blot Turbo Midi 0.2 µm PVDF Transfer Packs, pkg of 101704158	Trans-Blot Turbo Mini 0.2 µm Nitrocellulose Transfer Packs, pkg of 101704159
Trans-Blot Turbo Midi 0.2 µm Nitrocellulose Transfer Packs, pkg of 101704272	Trans-Blot Turbo RTA Mini 0.2 µm PVDF Transfer Kit, for 40 blots
1704273	Trans-Blot Turbo RTA Midi 0.2 µm PVDF Transfer Kit, for 40 blots
1704274	Trans-Blot Turbo RTA Mini 0.45 µm LF PVDF Transfer Kit, for 40 blots
1704275	Trans-Blot Turbo RTA Midi 0.45 µm LF PVDF Transfer Kit, for 40 blots
1704270	Trans-Blot Turbo RTA Mini 0.2 µm Nitrocellulose Transfer Kit,
for 40 blots
1704271	Trans-Blot Turbo RTA Midi 0.2 µm Nitrocellulose Transfer Kit,
for 40 blots
Imaging Systems
12009077 GelDoc Go Gel Imaging System with Image Lab Touch Software
12003153	ChemiDoc Imaging System12003154	ChemiDoc MP Imaging System
12018025 ChemiDoc Go Imaging System
Detection Reagents
1705060	C larity Western ECL Substrate, 200 ml
1705061	C larity Western ECL Substrate, 500 ml
1705062 Clarity Max Western ECL Substrate, 100 ml
12004158 StarBright Blue 700 Goat Anti-Mouse IgG, 400 µl
12004161 StarBright Blue 700 Goat Anti-Rabbit IgG, 400 µl
12005866 StarBright Blue 520 Goat Anti-Mouse IgG, 400 µl
12005869 StarBright Blue 520 Goat Anti-Rabbit IgG, 400 µl
12004163 hFAB Rhodamine Anti-Actin Primary Antibody, 200 µl
12004165 hFAB Rhodamine Anti-Tubulin Primary Antibody, 200 µl
12004167 hFAB Rhodamine Anti-GAPDH Primary Antibody, 200 µl

Bulletin 6427 Ver G US/EGTGX Stain-Free Precast Gels		Description

10-Well
50 μl

12-Well
20 μl

15-Well
15 μl

IPG Well
7 cm IPG Strip

Mini-PROTEAN TGX Stain-Free Precast Gels
7.5% Resolving Gel
45680234568024
10% Resolving Gel
45680334568034
12% Resolving Gel
45680434568044
4–15% Resolving Gel
45680834568084
4–20% Resolving Gel
45680934568094
8–16% Resolving Gel
45681034568104
Any kD Resolving Gel
456812345681244568025456803545680454568085456809545681054568125456802645680364568046456808645680964568106
4568126456802145680314568041456808145680914568101
4568121		
Description

10-Well
30 μl

12+2-Well*
45 μl

Criterion TGX Stain-Free Precast Gels**
7.5% Gel		
5678023
10% Gel		
5678033
12% Gel		
5678043
4–15% Gel		
5678083
4–20% Gel		
5678093
8–16% Linear Gradient5678103
Any kD Gel		
5678123

18-Well
30 μl26-Well
15 μl

Prep+2-Well*
700 μl

IPG+1-Well*
11 cm IPG Strip

56780245678034
56780445678084
56780945678104
567812456780255678035
56780455678085
567809556781055678125———5678082567809256781025678122———567808156780915678101
5678121* Reference wells accommodate 15 µl of markers/standards.
** Criterion TGX Stain-Free Gels are sold singly.

Visit bio-rad.com/StainFree for more information.
BIO-RAD, CHEMIDOC, CLARITY, MINI-PROTEAN, PRECISIONAB, STARBRIGHT, and TRANS-BLOT aretrademarks of Bio-Rad Laboratories, Inc. in certain jurisdictions.
TGX Stain-Free Precast Gels are covered by U.S. Patent Numbers 7,569,130 and 8,007,646.
Clarity Max Western ECL Substrate is manufactured for Bio-Rad by Cyanagen Srl and is the subject ofU.S. and ex-U.S. patents, including US7855287, EP1950207, US9040252, AU2011202658, CA2742025,
US8129136, EP1962095, and CN102313732.
All trademarks used herein are the property of their respective owner. © 2024 Bio-Rad Laboratories, Inc.

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