ゲルでタンパク質を染色

Published 7/08/2008
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Biology
 

Summary

電気泳動法で分離後、ゲル中のタンパク質は、いくつかの染色法で検出することができます。クマシーブルー、銀染色、SYPROオレンジ、SYPRO Rubyによるタンパク質の染色がこのビデオで実証されています。

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

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Abstract

電気泳動法で分離後、ゲル中のタンパク質は、いくつかの染色法で検出することができます。このユニットは、4つの一般的な方法によりタンパク質を検出するためのプロトコルについて説明します。クマシーブルー染色では、簡単かつ迅速な方法です。銀染色、より多くの時間がかかるが、はるかに敏感であるため、タンパク質の少量を検出するために使用することができますが。蛍光染色はクマシー染色よりも感度が主な理由は、伝統的な染色の手順に代わる空港として人気があります、そして多くの場合、銀染色と同様に敏感です。 SYPROオレンジとSYPROルビーを持つタンパク質の染色はまたここに実証されています。

Protocol

この実験的なアプローチのための完全なテキストのプロトコルでは使用可能です分子生物学におけるカレントプロトコール

Comments

8 Comments

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

    Reply
    Posted by: Anonymous
    August 3, 2008 - 7:45 AM
  2. thank you!very good!

    Reply
    Posted by: Anonymous
    September 6, 2008 - 8:51 PM
  3. It is very good, thanks

    Reply
    Posted by: Anonymous
    November 12, 2008 - 6:43 AM
  4. 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  

    Reply
    Posted by: Anonymous
    February 16, 2009 - 1:20 PM
  5. Itz realy ² gud!

    Reply
    Posted by: sathya r.
    May 18, 2009 - 11:01 AM
  6. Thank yoy

    Reply
    Posted by: JASSIM A.
    May 20, 2009 - 9:39 AM
  7. thank you

    Reply
    Posted by: ibrahim b.
    August 27, 2010 - 12:31 AM
  8. thanks! This is very useful for my job!

    Reply
    Posted by: Anonymous
    August 31, 2010 - 4:33 AM

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