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Séparation électrophorétique de protéines

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Proteomic Center, Keck Graduate Institute of Applied Life Sciences

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Cite this Article: Séparation électrophorétique de protéines

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

Abstract: Séparation électrophorétique de protéines

L'électrophorèse est utilisée pour séparer des mélanges complexes de protéines (par exemple, à partir des cellules, fractions subcellulaires, fractions de colonne, ou immunoprécipités), pour enquêter sur des compositions sous-unité, et de vérifier l'homogénéité des échantillons de protéines. Il peut également servir à purifier des protéines pour des applications supplémentaires. En électrophorèse sur gel de polyacrylamide, les protéines migrent en réponse à un champ électrique à travers les pores dans une matrice de gel de polyacrylamide, la taille des pores décroît avec la concentration d'acrylamide augmente. La combinaison de la taille des pores et chargé en protéines, la taille et la forme détermine le taux de migration de la protéine. Dans cette unité, la méthode standard Laemmli est décrite pour l'électrophorèse sur gel discontinu dans des conditions dénaturantes, c'est à dire, en présence de dodécylsulfate de sodium (SDS).

Protocol: Séparation électrophorétique de protéines

Le protocole texte complet de cette approche expérimentale est disponible en Current Protocols in Molecular Biology .

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Disclosures: Séparation électrophorétique de protéines

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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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