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Staining Proteins in Gels

1, 2

1Keck Graduate Institute of Applied Life Sciences, UVP, LLC, 2Proteomic Center, Keck Graduate Institute of Applied Life Sciences

 

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Cite this Article: Staining Proteins in Gels

Gallagher, S., Chakavarti, D. Staining Proteins in Gels. J. Vis. Exp. (17), e760, doi:10.3791/760 (2008).

Abstract: Staining Proteins in Gels

Following separation by electrophoretic methods, proteins in a gel can be detected by several staining methods. This unit describes protocols for detecting proteins by four popular methods. Coomassie blue staining is an easy and rapid method. Silver staining, while more time consuming, is considerably more sensitive and can thus be used to detect smaller amounts of protein. Fluorescent staining is a popular alternative to traditional staining procedures, mainly because it is more sensitive than Coomassie staining, and is often as sensitive as silver staining. Staining of proteins with SYPRO Orange and SYPRO Ruby are also demonstrated here.

Protocol: Staining Proteins in Gels

The complete text protocol for this experimental approach is available in Current Protocols in Molecular Biology.

Disclosures: Staining Proteins in Gels

The authors have nothing to disclose.

Ask the Author: Staining Proteins in Gels

8 Comments

WHICH REACTION EQUATION OF CONVERT Cu+2 TO Cu+1 in the protein test

1

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Posted by: najahAugust 3, 2008, 7:45 AM

thank you!very good!

2

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Posted by: yushaSeptember 6, 2008, 8:51 PM

It is very good, thanks

4

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Posted by: AnonymousNovember 12, 2008, 6:43 AM

A good and cheap alternative to SYPRO Rubi is RuBPS staining. Here are some protocols. More information can be found on www.ruthenium.ag.vu

 

RuBPS staining protocol I (quality)

1. Fix the gel in 30% EtOH, 10% acetic acid overnight

2. Rinse the gel in 20% EtOH for 30 min and repeat 3 times

3. Incubate the gel in 1 mM RuBPS solution for 6 h

4. Equilibrate the gel in water for 10 min and repeat once

5. Destain the gel with 40% EtOH/10% acetic acid for 15 h

6. Equilibrate the gel in water for 10 min repeat once and scan

 

all% are in V/V

 

Procedure as published in Proteomics 2004, 4, 599–608.

 

 

RuBPS staining protocol II (fast)

 

1. Incubate the gel in 50 ml of 40% Ethanol/10% acetic acid containing

   1 mM RuBPS for 1 h.

2. Destain the gel for 20 min in 40% Ethanol/10% acetic acid

3. Wash the gel for 10 min in water and scan

 

all% are in V/V

 

Procedure as published in PLoSONE. 2007 Feb 28;2(2)e263.

 

 

 

RuBPS staining protocol III (co-electrophoretical)

 

1. Add 1 ml of 20 mM stock solution to one pocket of your SDS gel. Run

    the gel according to your standard procedure.

2. Incubate the gel in 50 ml of 40% Ethanol/10% acetic acid for 20 min

3. Incubate the gel in 50 ml water for 10 min and scan

 

 

 

RuBPS staining protocol IV (loading buffer)

 

1. Add 1 ml of 20 mM stock solution to your loading buffer (omit all other

    dyes like bromophenol blue). Run the gel according to your standard

    procedure.

2. Incubate the gel in 50 ml of 40% Ethanol/10% acetic acid for 20 min

3. Incubate the gel in 50 ml water for 10 min and scan

 

5

Reply

Posted by: LamandaFebruary 16, 2009, 1:20 PM

Itz realy 2 gud!

7

Reply

Posted by: AnonymousMay 18, 2009, 11:01 AM

Thank yoy

12

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Posted by: JASSIM A.May 20, 2009, 9:39 AM

thank you

13

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Posted by: ibrahim b.August 27, 2010, 12:31 AM

thanks! This is very useful for my job!

14

Reply

Posted by: RichardTrinhAugust 31, 2010, 4:33 AM

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