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prepare sample protein for 2d from E.coli


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#1 somi

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Posted 07 December 2009 - 06:46 PM

I am doing 2D gel electrophoresis for E.coli. some one know what is the best method for preparing protein sample from E.coli .and is there any one know about how i can desalting my sample without use ready kit?

Edited by somi, 07 December 2009 - 06:47 PM.


#2 braincow

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Posted 10 December 2009 - 07:53 AM

You can find standard protocols just about anywhere. The Amersham 2D Electrophoresis handbook (pdf) has pretty good information on running 2D gels with bacterial proteins.

For desalting, I usually use the 2D cleanup kit from GE, which is widely used. If you prefer, you could google "TCA/acetone precipitation" and find a protocol, as the cleanup kit is basically this.

#3 somi

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Posted 10 December 2009 - 10:29 PM

You can find standard protocols just about anywhere. The Amersham 2D Electrophoresis handbook (pdf) has pretty good information on running 2D gels with bacterial proteins.

For desalting, I usually use the 2D cleanup kit from GE, which is widely used. If you prefer, you could google "TCA/acetone precipitation" and find a protocol, as the cleanup kit is basically this.



Thanks ,the handbook was very useful and i will try to TCA.




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