protocol for membrane receptor isolation

Kyle Legate via methods%40net.bio.net (by legatek from hotmail.com)
Fri Jun 27 14:00:46 EST 2008


WS wrote:
> Hi Engelbert,
> 
> a very nice protocol! It just came right in time as I need to do
> similar stuff: I have an antibody against a putative membrane
> receptor, so I want to incubate cells first with antobody at 4degC,
> then isolate membranes and then pick the antibody, hopefully with a
> tree of receptor and downstram proteins attached...
> 
> Just wonder if membrane proteins get solubilized without detegent?
> Will eg 0.5% TX100 disrupt the antibody-recptor complex?
> 
It shouldn't. My standard IP buffer in the past contained 1% TX 100.


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