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Elektrofores separation av proteiner

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Proteomic Center, Keck Graduate Institute of Applied Life Sciences

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Cite this Article: Elektrofores separation av proteiner

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

Abstract: Elektrofores separation av proteiner

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: Elektrofores separation av proteiner

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

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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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