0
I am new to the field of IHC and so am really hoping for some expert advice. I am looking for the protein gamma-h2ax which gets phosphorylated in the nucleus on double strand break formation and its a fairly common technique in the field of DNA repair. So am using a fixation method with ice-cold methanol and I also tried one with ethanol and acetic acid (which the Antibody company recommended). I felt the methanol looked better. However my gamma-h2ax foci don't look so clear and am not sure what I am doing wrong. I am right now thinking maybe of using a higher concentration of primary antibody (means a lower dilution). I am also putting up a representative image of my foci and hope somebody can look at it and tell me what I should be doing. Also does anyone think if using a detergeant like saponin might help.
DO HELP OUT... THANKS
