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Immunoprecipitation in Non-reducing conditions? - Protocol suggestions? (Oct/04/2006 )

Is there was a way to do an immunoprecipitation under non-reducing conditions? My protocols have always involved boiling the beads in SDS-PAGE sample buffer with DTT, but was hoping we could liberate our antigen in some way without using the reducing agent. I would greatly appreciate any information you could supply!

-lil-

you may be able to release the antigen by changing the pH to 2.5 or 9.5 to 10 as you would for immunoaffinity chromatography.

-mdfenko-

Boiling the sample without DTT will separate the anitgen from the antibody, and the antibody from the beads.

The binding is non-covalent so SDS alone should destroy any bond formation.

-jaknight-