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Die Färbung Proteine ​​in Gelen

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: Die Färbung Proteine ​​in Gelen

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

Abstract: Die Färbung Proteine ​​in Gelen

Nach der Trennung durch elektrophoretische Methoden können Proteine ​​in einem Gel durch mehrere Färbemethoden nachgewiesen werden. Diese Einheit beschreibt Protokollen zum Nachweis von Proteinen durch vier beliebtesten Methoden. Coomassie Blau-Färbung ist eine einfache und schnelle Methode. Silberfärbung, während mehr Zeit in Anspruch, ist wesentlich empfindlicher und können daher verwendet werden, um kleinere Mengen an Protein zu detektieren. Fluoreszenzfärbung ist eine beliebte Alternative zu den herkömmlichen Färbeverfahren, vor allem weil es empfindlicher als Coomassie-Färbung ist, und ist oft so empfindlich wie Silberfärbung. Die Färbung von Proteinen mit SYPRO Orange und SYPRO Ruby sind auch hier gezeigt.

Protocol: Die Färbung Proteine ​​in Gelen

Der vollständige Text Protokoll für diesen experimentellen Ansatz ist in Current Protocols in Molecular Biology .

Disclosures: Die Färbung Proteine ​​in Gelen

Ask the Author: Die Färbung Proteine ​​in Gelen

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

Reply

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

thank you

13

Reply

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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