Elektrofores separation av proteiner

Biology

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Summary

I denna video visar vi en metod för elektrofores separation av proteiner med hjälp av poly-acrylimide gelelektrofores (sida).

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

Elektrofores används för att separera komplexa blandningar av proteiner (t.ex. från celler, subcellulära bråk, bråk kolumn eller immunoprecipitates), för att undersöka underenhet kompositioner, och att bekräfta homogenitet av protein prover. Den kan också användas för att rena proteiner för användning i ytterligare ansökningar. I polyakrylamidgelelektrofores, proteiner vandrar som svar på ett elektriskt fält genom porer i en polyakrylamidgel matris, porstorlek minskar med ökande akrylamid koncentration. Kombinationen av porstorlek och protein laddning, storlek och form bestämmer migration graden av proteinet. I denna enhet, är standard Laemmli beskrivna metoden för diskontinuerliga gel elektrofores under denaturering villkor, det vill säga i närvaro av natriumdodecylsulfat (SDS).

Protocol

Den fullständiga texten protokollet för denna experimentella strategi finns i nuvarande protokoll i molekylärbiologi .

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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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