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Separazione elettroforetica delle proteine

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Proteomic Center, Keck Graduate Institute of Applied Life Sciences

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Cite this Article: Separazione elettroforetica delle proteine

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

Abstract: Separazione elettroforetica delle proteine

Elettroforesi viene utilizzata per separare miscele complesse di proteine ​​(per esempio, dalle cellule, frazioni subcellulari, frazioni colonna o immunoprecipitati), per indagare composizioni subunità, e per verificare l'omogeneità dei campioni di proteine. Può anche servire a purificare le proteine ​​per l'uso in altre applicazioni. Elettroforesi in gel di poliacrilamide, proteine ​​migrano in risposta ad un campo elettrico attraverso i pori in una matrice di gel di poliacrilammide; dimensione dei pori diminuisce con l'aumentare la concentrazione di acrilamide. La combinazione di dimensione dei pori e la carica di proteine, le dimensioni e la forma determina il tasso di migrazione della proteina. In questa unità, il metodo standard Laemmli è descritto per elettroforesi su gel discontinuo in condizioni di denaturazione, cioè, in presenza di sodio dodecil solfato (SDS).

Protocol: Separazione elettroforetica delle proteine

Il protocollo testo completo di questo approccio sperimentale è disponibile in protocolli di corrente in Biologia Molecolare .

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Disclosures: Separazione elettroforetica delle proteine

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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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