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Electrophoretic Separation of Proteins

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Proteomic Center, Keck Graduate Institute of Applied Life Sciences

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Cite this Article: Electrophoretic Separation of Proteins

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

Abstract: Electrophoretic Separation of Proteins

Electrophoresis is used to separate complex mixtures of proteins (e.g., from cells, subcellular fractions, column fractions, or immunoprecipitates), to investigate subunit compositions, and to verify homogeneity of protein samples. It can also serve to purify proteins for use in further applications. In polyacrylamide gel electrophoresis, proteins migrate in response to an electrical field through pores in a polyacrylamide gel matrix; pore size decreases with increasing acrylamide concentration. The combination of pore size and protein charge, size, and shape determines the migration rate of the protein. In this unit, the standard Laemmli method is described for discontinuous gel electrophoresis under denaturing conditions, i.e., in the presence of sodium dodecyl sulfate (SDS).

Protocol: Electrophoretic Separation of Proteins

The complete text protocol for this experimental approach is available in Current Protocols in Molecular Biology.

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Disclosures: Electrophoretic Separation of Proteins

The authors have nothing to disclose.

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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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