western blotting and histone acetylation - (Aug/21/2005 )

hi!
im trying to do western analysis with antiacetyl histone4 antibody. the protein is a small one: ~11kDA. i dont know why, but i cant see any proteins smaller then 25 kDA...they just gone! maybe the small proteins are running out of the membrane while blotting...i dont know!
if some one have an experience with w.b. to see histones, please tell me how you are doing this!

+ do you think i should take only nuclear proteins or maybe its o'k to use the whole cell lysate?


thanks a lot!!!!

nuclear lysate will certainly be alot cleaner,

are you acid extracting your proteins, as you know histones are within a complex and form the nucleosome, acid extraction will disssociate the histones and you should then see them.

I ain't in the office o give you a protocol but if you google it I am sure it will come up!

Nick