タンパク質の電気泳動分離

Biology

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Summary

このビデオでは、我々は、ポリ- acrylimideゲル電気泳動(PAGE)を用いてタンパク質の電気泳動分離する方法を示しています。

Cite this Article

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Chakavarti, B., Chakavarti, D. Electrophoretic Separation of Proteins. J. Vis. Exp. (16), e758, doi:10.3791/758 (2008).

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Abstract

電気泳動はタンパク質の複雑な混合物を分離する(例えば、細胞、細胞内の分画、カラム画分、または免疫沈降から)、サブユニットの組成を調査するため、およびタンパク質サンプルの均質性を検証するために使用されます。また、さらにアプリケーションで使用するためのタンパク質を精製するのに役立つことができる。ポリアクリルアミドゲル電気泳動では、蛋白質はポリアクリルアミドゲルマトリックスの細孔を通して電界に反応して移行し、細孔サイズが増加するアクリルアミドの濃度が減少する。細孔径と蛋白質の電荷、大きさ、および形状の組み合わせは、タンパク質の移行速度を決定します。このユニットでは、標準をLaemmli法は、ドデシル硫酸ナトリウム(SDS)の存在下で、変性条件下で不連続ゲル電気泳動のために、すなわち記述されている。

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

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Comments

10 Comments

  1. Hi nice demo but I did not understand why the presenters kept on showing an agarose gel electrophoresed DNA gel in between as a representation instead of a protein gel.

    Reply
    Posted by: Anonymous
    June 15, 2008 - 1:11 PM
  2. SYPROorange and SYPROruby staining - two fluorescent-based protein detection methods - can appear like ethidium bromide stained DNA gels.  So those gels you are speaking of may actually be protein gels. 

    Reply
    Posted by: Anonymous
    June 16, 2008 - 9:34 AM
  3. Yeah possible,  but then why do we see only a single band.I guess SYPRO detection methods would detect the entire protein panel and not specific bands. unless otherwise they are detecting some specific enzyme activity using flourescent detection methods.

    Reply
    Posted by: Anonymous
    June 16, 2008 - 9:52 AM
  4. We loaded BSA.

    Thanks.

    Reply
    Posted by: Anonymous
    June 17, 2008 - 2:12 AM
  5. Please not that we showed polyacrylamide gel electrophoresis and not agarose gel electrophoresis.

    Thanks.

    Bulbul Chakravarti

    Reply
    Posted by: Anonymous
    June 17, 2008 - 2:08 AM
  6. why cannot i see the video...? pls help me..

    Reply
    Posted by: Anonymous
    October 10, 2008 - 3:35 AM
  7. i can see it now..thanks...

    Reply
    Posted by: Anonymous
    October 10, 2008 - 4:50 AM
  8. my network is poor, I can not enjoy the video

    Reply
    Posted by: Huang C.
    March 15, 2009 - 6:30 AM
  9. Hello, Please shoot me an email at nikitab@jove.com and we'll figure something out for you. Cheers, Nikita

    Reply
    Posted by: Anonymous
    March 22, 2009 - 10:26 PM
  10. Hi, thanks what can I download your video ? please help me

    Reply
    Posted by: violet s.
    January 14, 2012 - 10:35 AM

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