How to elute antigens without eluting Ab off Prot. A?

Noam Harel Harel_n at a1.mscf.upenn.edu
Sun Dec 17 21:53:13 EST 1995


Hi,

	I am performing immunoprecipitations on unlabeled nuclear
extracts using non-covalently linked Protein A-antibody combinations.
The problem is, when I run the precipitates on SDS-PAGE, then do a 
Western blot, the secondary antibody on the W. blot causes the heavy
 chains of the Abs used in the ip to create a 
huge smear that covers up many of the potential bands I'd like to see.
Is there any way besides covalently linking the Ab to the beads, 
or besides eluting with competitor peptides, to keep the Ab from
coming off the Prot. A beads when boiling in SDS-PAGE buffer?
I heard something about elution using glycine, but can't find a protocol
Thanks for anyhelp,

N. Harel




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