Separación electroforética de las proteínas

Biology

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Summary

En este vídeo, se demuestra un método para la separación electroforética de las proteínas usando poli-acrylimide electroforesis en gel (PAGE).

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

Electroforesis se utiliza para separar mezclas complejas de proteínas (por ejemplo, a partir de células, fracciones subcelulares, las fracciones de la columna, o inmunoprecipitados), para investigar las composiciones de subunidades, y para verificar la homogeneidad de las muestras de proteínas. También puede servir para purificar las proteínas para su uso en otras aplicaciones. En la electroforesis en gel de poliacrilamida, las proteínas migran en respuesta a un campo eléctrico a través de los poros de una matriz de gel de poliacrilamida; tamaño de poro disminuye con la concentración de acrilamida en aumento. La combinación de tamaño de poro y la carga de proteínas, el tamaño y forma que determina la tasa de migración de la proteína. En esta unidad, el método estándar de Laemmli se describe, por electroforesis en gel discontinuo bajo condiciones de desnaturalización, es decir, en presencia de dodecil sulfato sódico (SDS).

Protocol

El protocolo de texto completo de este enfoque experimental está disponible en Current Protocols in Molecular Biology .

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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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