co-ip of bacteria membrane protein

[Dr Engelbert Buxbaum]

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.