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젤의 단백질 염색법

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: 젤의 단백질 염색법

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

Abstract: 젤의 단백질 염색법

전기 방법으로 분리에 따라, 젤의 단백질은 몇 얼룩 방법에 의해 감지하실 수 있습니다. 이 장치 네 인기가 방법으로 단백질을 검출 프로토콜을 설명합니다. 쿠매시 푸른 얼룩이 간단하고 빠른 방법입니다. 실버 얼룩, 더 많은 시간을 소모는 상당히 민감한이며, 따라서 단백질의 작은 금액을 감지하는 데 사용할 수있는 반면. 그것이 쿠매시의 얼룩보다 더 민감하고, 종종 실버 얼룩처럼 민감 주로하기 때문에 형광 염색법은 전통적인 염색법 절차에 인기있는 대안입니다. SYPRO 오렌지 SYPRO 루비와 단백질 얼룩도 여기에 증명하고 있습니다.

Protocol: 젤의 단백질 염색법

이 실험 방법에 대한 전체 텍스트 프로토콜에서 사용할 수 있습니다 분자 생물학의 현재 프로토콜 .

Disclosures: 젤의 단백질 염색법

Ask the Author: 젤의 단백질 염색법

8 Comments

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

1

Reply

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

thank you!very good!

2

Reply

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

It is very good, thanks

4

Reply

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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