2 replies to this topic

[flyhead]

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!

[Dave_Kub_11]

flyhead, on 17 February 2011 - 01:42 PM, said:

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!

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

[castleinthesky]

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