Coomassie Blue Gel and Membrane Stains | Thermo Fisher Scientific

The most common method for in-gel protein detection is staining with Coomassie blue dye. Coomassie dye staining is especially convenient because it involves a single, ready-to-use reagent and does not permanently chemically modify the target proteins. Because no chemical modification occurs, excised protein bands can be completely de-stained and the proteins recovered for analysis by mass spectrometry or sequencing. We offer several Coomassie based protein stains that are easy-to-use and provides a simple visualization of proteins resolved by electrophoresis.

Compare different protein staining methods

Highlights of Coomassie stains:

Easy-to-use—simple protocols with only 2-3 steps
Compatible—Coomassie based stains are compatible with many downstream applications such as quantitative densitometry, mass spectrometry or sequencing analysis
Reversible

Find the right Coomassie based stain for your research needs

		Simply Blue Safe Stain	Colloidal Blue Staining	GelCode Blue Stain	GelCode Blue Safe Stain	Imperial Protein	Pageblue Protein Stain
Detection limit		> 7 ng	< 10 ng	8 ng	9 ng	≤3 ng	5 ng
Approx. staining time (for gels)	Std	135 min	10 hr	135 min	135 min	135 min	95 min
Approx. staining time (for gels)	Microwave	12 min		30 min	30 min	12 min	30 min
# Components		1 (Ready-to-use)	2	1 (Ready-to-use)	2	1 (Ready-to-use)	1 (Ready-to-use)
# of staining steps		2	3	2-3	3	3	3
Staining agent		G-250	G-250	G-250	G-250	R-250	G-250
Highlights		Non-hazardous disposal		Flexible stain for multiple applications	Non-hazardous shipping	Purple stain	Stain can be reused up to 3 times
No acid fixative or methanol required		✓	Methanol is required for staining, and Bis-tris gels require fixing prior to staining	✓ *Bis-tris gels require fixing prior to staining	✓	✓	✓
Gel staining		✓	✓	✓	✓	✓	✓
PVDF membrane staining		✓ *membrane must be dry	×	✓	✓	×	✓ *membrane must be dry
Nitrocellulose membrane staining		×	×	✓	✓	✓	×
MS or sequencing analysis		✓	✓	✓	✓	✓	✓
Catalog number		LC6065	LC6025	24592	24594	25615	24620

Coomassie Staining Protocol

An initial water wash step is necessary to remove residual SDS, which interferes with dye binding. Then the staining reagent is added, usually for about 1 hour with rocking. The gel can be analyzed directly in the stain. Alternatively, a water or methanol:acetic acid de-staining step can be used to wash away excess unbound dye from the gel matrix. To expedite the staining process microwaves can be used with Coomassie stains not requiring methanol or acetic acid.

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SimpleBlue SafeStain protocol

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Imperial Protein Stain Protocol

Sensitivity

3-10 microgram of protein can be detected usually within 5 minutes in Coomassie stains. With additional water-based de-staining, as little as 7 ng of protein (BSA) can be detected.

Lane 1: 6 µg protein mix
Lane 2: 1 µg rabbit IgG
Lane 3: 1 µg reduced BSA
Lane 4: 5 µg E. coli lysate
Lane 5: 20 ng reduced BSA
Lane 6: 10 ng reduced BSA
Lane 7: 7 ng reduced BSA
Lane 8: 3 ng reduced BSA
Lane 9: 10 µl Mark12 Standard
Lane 10: 5 µl Mark12 Standard

NuPAGE Bis-tris gel stained with SimplyBlue SafeStain for 1 hour and destained overnight in water.

Rapid staining with Imperial protein stain

Protein bands can be detected after 5 minutes with staining with Imperial Protein Stain. For greater sensitivity and reduced background, gels can be stained for 1 hour and de-stained overnight in water.

Linearity

Coomassie blue dyes bind proteins quantitatively within a certain protein range allowing for densitometry analysis. PageBlue protein stain can deliver a dynamic range of ~5ng to ~500ng.

Linear dynamic range of PageBlue Protein Stain

PageBlue Protein Staining Solution is sensitive and has a broad, linear dynamic range. A series of samples containing beta-galactosidase and bovine serum albumin (0 to 500ng each) were separated by electrophoresis on a 12% Tris-glycine SDS-PAGE, gel, and then stained with PageBlue Protein Staining Solution and densitometry was performed on the stained protein bands.

Capture and analysis of Coomassie stained gels and membranes

iBright Imaging Systems

Capture and analyze publication quality images from stained gels and membranes with iBright Imaging Systems. These high-performance instruments enhance the imaging experience through powerful hardware, advanced automated technologies, and an interface that is easy to use for researchers of all experience levels.

Explore our iBright gel documentation systems

Dual LED Blue/White Light Transilluminator

Illuminate your stained gels and membranes with this white light box.

Learn more about the Dual LED Transilluminator

Overview

Find the right Coomassie based stain for your research needs

		Simply Blue Safe Stain	Colloidal Blue Staining	GelCode Blue Stain	GelCode Blue Safe Stain	Imperial Protein	Pageblue Protein Stain
Detection limit		> 7 ng	< 10 ng	8 ng	9 ng	≤3 ng	5 ng
Approx. staining time (for gels)	Std	135 min	10 hr	135 min	135 min	135 min	95 min
Approx. staining time (for gels)	Microwave	12 min		30 min	30 min	12 min	30 min
# Components		1 (Ready-to-use)	2	1 (Ready-to-use)	2	1 (Ready-to-use)	1 (Ready-to-use)
# of staining steps		2	3	2-3	3	3	3
Staining agent		G-250	G-250	G-250	G-250	R-250	G-250
Highlights		Non-hazardous disposal		Flexible stain for multiple applications	Non-hazardous shipping	Purple stain	Stain can be reused up to 3 times
No acid fixative or methanol required		✓	Methanol is required for staining, and Bis-tris gels require fixing prior to staining	✓ *Bis-tris gels require fixing prior to staining	✓	✓	✓
Gel staining		✓	✓	✓	✓	✓	✓
PVDF membrane staining		✓ *membrane must be dry	×	✓	✓	×	✓ *membrane must be dry
Nitrocellulose membrane staining		×	×	✓	✓	✓	×
MS or sequencing analysis		✓	✓	✓	✓	✓	✓
Catalog number		LC6065	LC6025	24592	24594	25615	24620

How it works

Coomassie Staining Protocol

Click image to enlarge

SimpleBlue SafeStain protocol

Click image to enlarge

Imperial Protein Stain Protocol

Sensitivity

3-10 microgram of protein can be detected usually within 5 minutes in Coomassie stains. With additional water-based de-staining, as little as 7 ng of protein (BSA) can be detected.

NuPAGE Bis-tris gel stained with SimplyBlue SafeStain for 1 hour and destained overnight in water.

Linearity

Coomassie blue dyes bind proteins quantitatively within a certain protein range allowing for densitometry analysis. PageBlue protein stain can deliver a dynamic range of ~5ng to ~500ng.

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