Detergent Concentrations for Co-IP - (Sep/06/2005 )

Does anyone know what is an appropriate detergent concentration for detection of interacting proteins on Co-IP? I'm using 1% NP-40 now and cannot reproduce previously reported interactions although the IP's work fine.

Should I try a milder and/or less stringent detergent solution? Has anyone tried CHAPS, OPG, or Triton-X before?

Thanks

-Mountainman-

QUOTE (Mountainman @ Sep 6 2005, 07:08 PM)

I use this IP lysis buffer for my IPs. Works well for me:

IP Lysis Buffer
20mM HEPES pH 7.4
150mM NaCl
0.5% NP-40
1mM EDTA
1mM EGTA
10% Glycerol

Hope this helps.

LTR

-L_Reiter-

try to go down with NP40. also salt concentration is important. 150mM NaCl is considered to be mild.

-Jusu-