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Elektrophoretische Trennung von Proteinen

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Proteomic Center, Keck Graduate Institute of Applied Life Sciences

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Cite this Article: Elektrophoretische Trennung von Proteinen

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

Abstract: Elektrophoretische Trennung von Proteinen

Elektrophorese wird verwendet, um komplexe Gemische von Proteinen (z. B. von Zellen, subzellulären Fraktionen, Säulenfraktionen oder Immunpräzipitaten) zu trennen, die Untereinheit Kompositionen zu untersuchen und um die Homogenität der Protein-Proben zu überprüfen. Es kann auch dazu dienen, um Proteine ​​für den Einsatz in weiteren Anwendungen zu reinigen. In Polyacrylamidgelelektrophorese, Proteine ​​in Reaktion auf ein elektrisches Feld durch die Poren in einem Polyacrylamid-Gel-Matrix zu migrieren, Porengröße nimmt mit steigender Acrylamid-Konzentration. Die Kombination von Porengröße und Protein Ladung, Größe und Form bestimmt die Wanderungsgeschwindigkeit des Proteins. In dieser Einheit ist die Standard-Laemmli-Methode für die diskontinuierliche Gelelektrophorese unter denaturierenden Bedingungen beschrieben, dh in Gegenwart von Natriumdodecylsulfat (SDS).

Protocol: Elektrophoretische Trennung von Proteinen

Der vollständige Text Protokoll für diesen experimentellen Ansatz ist in Current Protocols in Molecular Biology .

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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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