cell fractionation plus IP - (Sep/27/2005 )
QUOTE (miguelon @ Oct 10 2005, 01:04 PM)
Sorry, maybe had not read properly your reply, you're suggesting to add the detergent after the fractionation step. Is a good idea.
About the crosslinking, I have not tried by myself, I was asking you whether you knew something about it. I think that some chemical crosslinking approaches are reversible, like PFA. Maybe you could crosslink direct interactions in that fraction, in order to apply harsher solubilizing conditions and keep stable your target interaction during that step? Has anybody tried that approach? Is it only suitable for in vitro studies?
About the crosslinking, I have not tried by myself, I was asking you whether you knew something about it. I think that some chemical crosslinking approaches are reversible, like PFA. Maybe you could crosslink direct interactions in that fraction, in order to apply harsher solubilizing conditions and keep stable your target interaction during that step? Has anybody tried that approach? Is it only suitable for in vitro studies?
hmm.. never tried that.. and really have no idea..
-Jusu-
Have you ever tried a Fractionating Column? Some companies provide these, one that stands out is the ChromBA column.
Good luck, and never quit trying!
-Dr. Seninger-
By the way, it is also likely that Invitrogen or Qiagen has a kit for this. Not sure though...
Good luck!
-Dr. Seninger-