Protein cross-linking in crude bacterial cell extract.

Nikolai Chitaev nikolaic at VISAR.WUSTL.EDU
Fri Sep 27 17:23:17 EST 1996


> Date: Fri, 27 Sep 1996 16:25:58 -0400
 Bin LIN <binlin at umich.edu> wrote:
> Hello netters:
> 
> I am planning to indentify proteins, in crude bacterial cell extract,
> interacting with a bacterial G-protein by chemical cross-linking.
> 
> As you might know, there is 0.1mM PMSF in the cell extract buffer. I
> am wondering whether PMSF will interact with cross-linker (DMA or
> DMP).

It should not.

> Could anyone give me some advice on setting isotonic extracting buffer
> 
> and choosing appropriate cross-linkers? BTW, what's the best
> terminating buffer for cross-linking reaction?

1 M glycin, or 1M Tris. The x-linker works on primary amines.
There is such thing as PIERCE catalog. Look up there.
-- 
Nikolai
http://128.252.119.253



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