You could try the method used by Kane and colleagues years ago when looking at V-ATPase complexes in yeast. See Kane, PM (1995) JBC 270:17025-32, PMID: 7622524. She treated extracts with a reversible crosslinker, DSP, before the IP step.

Thanks a lot!!!!!

Pierce have some good resources for cross-linking. Lots of info here: http://www.piercenet.com/browse.cfm?fldID=020306

I did some crosslinking for IP a long time ago, and it was tricky to work out which crosslinker was appropriate. I found the Pierce resources very helpful.

Thanks Samantha!!!!

The procedure to cross-link between protein-protein and protein-DNA using formaldehyde are widely used in Chromatin-immunoprecipitation. The procedures are available established and online from Upstate, NEBioLab, Promega, Clontech, Pierce, QiaGen or many other relevant publications.

The reversal of cross-linking is performed by boiling in the presence of SDS. Analysis by PAGE and Western-blotting. I have used the identification of minute quantities of IP proteins using this procedures. Please refer to my recent paper in J.Biol.Chem. 2010.

If you use a 35S-methionine labelled protein you can visualize by autoradiography and you will eliminate the disturbance of IgGs used in the IP in your Western Blots.

Best of luck.

I have attached a Protocol Guide from ClonTech in PDF

Thanks a lot manjapra!!!!

an alternative to cross-linking is to covalently couple the Abs directly to the bead surface using epoxy or tosyl chemistry.

Nucleic Acids Research, 2014, Vol. 42, No. 14 9021