| | Re: Problems with Immunoprecipitation and Cross-linking
I guess if I checked this forum more often I could've replied earlier, I'm not sure if this is still troubling you.
Anyways, like kevin_a said, I was trying to do a similar pull-down experiment with dynabeads protein G. My cross linking reaction did work except that I couldn't get method of eluting just the protein. I tried Citrate ph 3 buffer in their manual which gave me no protein, however when I boiled the same beads in SDS loading buffer I saw the protein on the western blot gel.
However, I ended up forsaking the cross-linking because like Kevin said,if you can resolve the protein from the antibody heavy chain, you shouldn't need cross-linking. And in my case, I had a negative control antibody(preinjection serum) which I ran along that clearly told me that the additional band is the protein that I pulled down.
Last edited by marcroboy; 08-12-2008 at 02:22 AM.