电泳分离蛋白质

Biology

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Summary

在这段视频中,我们展示了使用acrylimide聚凝胶电泳(PAGE)蛋白电泳分离的方法。

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

电泳用于分离复杂的蛋白质混合物(例如,从细胞,亚细胞组分,列分数,或免疫),调查亚基组成,并验证蛋白质样品的同质性。 ,它也可以用来净化在进一步的应用中使用的蛋白质。在聚丙烯酰胺凝胶电泳,蛋白质通过毛孔中的聚丙烯酰胺凝胶基质中的电场响应迁移;孔径丙烯酰胺浓度的增加而降低。孔径和蛋白质的电荷,大小和形状的组合决定了蛋白质的迁移率。在这个单元,标准的Laemmli方法描述不连续的凝胶电泳变性条件下,也就是说,在十二烷基硫酸钠(SDS),的存在。

Protocol

这个实验方法的完整的文本协议,可用在分子生物学中的电流协议。

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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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