detect histone acetylation - (Feb/17/2011 )

I have recently done some Western blots with fruit flies. I have no problem in getting signal for histone 3 but have a hard time to detect any acetylated H3 even if I increase the amount of tissue for lysis. I saw some papers mentioned using HDAC inhibitors, such as sodium butyrate when lysing. Does anybody have experience or have suggestions? Thanks in advance!

flyhead on Thu Feb 17 21:42:49 2011 said:

May be off topic with what you are looking at but the only I've been able to detect ap Histone 3 is by enriching the protien by immunoprecipiatation. The trouble is the light chain of the secondary antibody appears roughly above where ap Histone 3 should appear on the blot. But I guess that depends on the antibody you are IP'ing with. Try the IP

HDAC inhibitors (HDI) can alter the degree of acetylation nonhistone proteins and be used in cancer treatment.