Gentle extraction of nuclear protein complex - (Apr/06/2008 )

I have been trying to isolate a nuclear protein complex but have had trouble getting any binding partners. So I am looking for a recipe or product that will extract nuclear proteins but is as mild as possible as to not disrupt any complexes. I have tried using M-PER from pierce, standard 1% NP-40 recipes (without DTT) and more stringent RIPA buffers. Any suggestions?

-reddpoint-

Have you tried something like crosslinking the proteins, disrupting the nucleus and then proceed to the IP?

If it works in ChIP methods, it should also work for straight IPs.

Hope this helps

-Madrius-

QUOTE (Madrius @ Apr 7 2008, 05:35 AM)

Have you tried something like crosslinking the proteins, disrupting the nucleus and then proceed to the IP?

If it works in ChIP methods, it should also work for straight IPs.

Hope this helps

I have done formaldehyde crosslinking ala CHIP which did yield some bands but I was worried about false positives.

-reddpoint-