co-ip of bacteria membrane protein

Dr Engelbert Buxbaum engelbert_buxbaum at hotmail.com
Tue Nov 4 00:33:45 EST 2003


mj wrote:

> Hi, all
> 
> I am planning to do a co-IP on an inner membrane protein with HA tag of
> Caulobacter(a gram negative bacterium), and I am hoping to pull down some
> protein complexes under non-denaturing condition (low detergent).  Has
> anybody done similar experiments like this?  I really don't know where to
> start, does anybody have a protocol for that?
> 
> How shall I determine the right condition that can solubilize the membrane
> protein yet maintain the complexes?  For HA tag, which antibody will be
> better, monoclonal or polyclonal?

Calbiochem offers heat-killed protein A (Pansorbin) or protein G
containing bacteria for immunoprecipitation experiments, and they have
published a little booklet on methods. You may wish to consult this. 

Usual disclaimer applies.



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