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Elektroforetische scheiding van eiwitten

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Proteomic Center, Keck Graduate Institute of Applied Life Sciences

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Cite this Article: Elektroforetische scheiding van eiwitten

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

Abstract: Elektroforetische scheiding van eiwitten

Elektroforese wordt gebruikt om complexe mengsels van eiwitten (bijvoorbeeld van cellen, subcellulaire fracties, kolom breuken of immunoprecipitaten) te scheiden, om subunit composities te onderzoeken en om de homogeniteit van eiwit monsters te controleren. Het kan ook dienen om eiwitten te zuiveren voor gebruik in verdere toepassingen. In polyacrylamidegelelektroforese, eiwitten migreren als reactie op een elektrisch veld door de poriƫn in een polyacrylamide gel matrix; poriegrootte afneemt met toenemende acrylamide concentratie. De combinatie van poriegrootte en eiwit lading, de grootte en de vorm bepaalt de migratie snelheid van het eiwit. In deze unit, is de standaard Laemmli methode beschreven voor discontinue gelelektroforese onder denaturerende omstandigheden, dat wil zeggen, in het bijzijn van natriumdodecylsulfaat (SDS).

Protocol: Elektroforetische scheiding van eiwitten

De volledige tekst protocol voor deze experimentele aanpak is beschikbaar in Current Protocols in Molecular Biology .

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Disclosures: Elektroforetische scheiding van eiwitten

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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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