Lysis of the cells - (Jul/01/2010 )
Dear ChIP-friends,
I am facing a big problem. The HeLa cells which I am using for ChIP do not lyse in SDS buffer anymore after they were infected with adenovirus. No lysis method used is able to break the cells, not even protein sample (laemmli-buffer). Has anyone ever faced this problem and could give me any useful hints?
Thank you
Nicole
-nipa68-
nipa68 on Jul 1 2010, 01:07 AM said:
Dear ChIP-friends,
I am facing a big problem. The HeLa cells which I am using for ChIP do not lyse in SDS buffer anymore after they were infected with adenovirus. No lysis method used is able to break the cells, not even protein sample (laemmli-buffer). Has anyone ever faced this problem and could give me any useful hints?
Thank you
Nicole
I am facing a big problem. The HeLa cells which I am using for ChIP do not lyse in SDS buffer anymore after they were infected with adenovirus. No lysis method used is able to break the cells, not even protein sample (laemmli-buffer). Has anyone ever faced this problem and could give me any useful hints?
Thank you
Nicole
Are you saying that they're not lysed because you see particulate matter after the lysis step? It's normal to see particulate matter after the lysis step if the cells have been crosslinked for ChIP since the nuclei and cytoskeleton remain intact throughout lysis. The only thing that disrupts the nuclei and cytoskeleton is the shearing step.
-KPDE-