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Separación electroforética de las proteínas

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Proteomic Center, Keck Graduate Institute of Applied Life Sciences

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Cite this Article: Separación electroforética de las proteínas

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

Abstract: Separación electroforética de las proteínas

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: Separación electroforética de las proteínas

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

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Disclosures: Separación electroforética de las proteínas

Ask the Author: Separación electroforética de las proteínas

5 Comments

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.

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Posted by: swethaJune 15, 2008, 1:11 PM

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. 

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Posted by: Aaron K.June 16, 2008, 9:34 AM

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.

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Posted by: swethaJune 16, 2008, 9:52 AM

We loaded BSA.

Thanks.

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Posted by: Bulbul ChakravartiJune 17, 2008, 2:12 AM

Please not that we showed polyacrylamide gel electrophoresis and not agarose gel electrophoresis.

Thanks.

Bulbul Chakravarti

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Posted by: Bulbul ChakravartiJune 17, 2008, 2:08 AM

why cannot i see the video...? pls help me..

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Posted by: AnonymousOctober 10, 2008, 3:35 AM

i can see it now..thanks...

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Posted by: AnonymousOctober 10, 2008, 4:50 AM

my network is poor, I can not enjoy the video

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Posted by: Huang C.March 15, 2009, 6:30 AM

Hello,

Please shoot me an email at nikitab@jove.com and we'll figure something out for you.

Cheers,

Nikita

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Posted by: AnonymousMarch 22, 2009, 10:26 PM

Hi, thanks what can I download your video ? please help me

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

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