Proteinlerin Elektrogöçer Ayırma

Biology

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Summary

Bu video, poli-acrylimide jel elektroforezi (PAGE) kullanarak proteinlerin elektroforetik ayrımı için bir yöntem ortaya koymaktadır.

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

Elektroforez, proteinlerin (örneğin, hücre, hücre içi kesirler, sütun kesirler, veya immunoprecipitates) kompleks karışımlar ayrı altbirim kompozisyonları araştırmak için, protein örneklerinin homojenliği ve doğrulamak için kullanılır. Ayrıca başka uygulamalar için protein arındırmak için hizmet edebilir. Poliakrilamid jel elektroforezi, proteinler poliakrilamid jel matris gözenekler yoluyla bir elektrik alanı yanıt göç; gözenek büyüklüğü artan akrilamid konsantrasyonu azalır. Gözenek boyutu ve protein yükü, boyut ve şekil kombinasyonu protein göç oranı belirler. Bu ünitede, standart Laemmli yöntemi, sodyum dodesil sülfat (SDS) varlığı, denatüre edici koşullar altında yani süreksiz jel elektroforezi için açıklanmıştır.

Protocol

Bu deneysel bir yaklaşım için tam metin protokolü mevcuttur Moleküler Biyoloji Mevcut Protokoller .

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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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