[Histonet] Alkaline Phospahtase and Methyl Green

[Igor Deyneko]

Dear Histonetters!
I was wondering if anyone can help me out. I'm doing IHC with Ach antibodies
and when I use DAB chromogen I get strong background. I used Vulcan Red
(from Biocare Medical), which got rid of the background, but it's a bit hard
to distinguish from Hematoxylin counterstain that I use. I was wondering if
Methyl Green would be an appropriate counterstain for it? I tried
using 0.5%Methyl Green from Plyscientific before, but had problems
because it washed
off right away in the first change of 95 Ethanol. I also tried n-butanol and
acetone, but the results were unsatisfactory. If anyone knows good
percentage to use as a counterstain and a protocol would be greatly
appreciated.
Thank you in advance.
Igor Deyneko
Infinity Pharmaceuticals
In-Vivo Pharmacology
Cambridge, MA