Eliminate light/heavy chain after IP??

Andrei Popov andrei.popov at bbsrc.ac.uk
Fri Aug 23 11:39:03 EST 1996


Danny Altschuler wrote:
> 
> I'm doing some IP/Western assays. My protein comigrate exactly with the
> light chain, so after IP and blotting the light chain mask the detection
> of my protein. I've tryied Laemli buffer without DTT/betaMSH and even
> though 95% of the IgG runs as a high MW, the remaining 5% still is enough
> to interfere with detection. Does anyone know any protocol to get rid of
> IgG after IPs?
> Please, e-mail me to
> altschul at med.unc.edu
> Thanks
> Danny


Hello,

I would suggest to use for detection of your primary antibodies
a heavy chain-specific conjugate. We use quite a lot of heavy- or light-chain
(either kappa or lambda) specific biotinilated antibodies and usually do not have 
background problems.

best wishes

Andrei



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