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prepare sample protein for 2d from E.coli - (Dec/07/2009 )

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?

-somi-

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.

-braincow-

braincow on Dec 10 2009, 11:53 PM said:

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.

-somi-