[Histonet] Need help with CD40 on frozen sections

[Bea DeBrosse-Serra]

Histonetters,

I need some help with CD40 on frozen sections (spleen, non-treated). I cut my frozens, let them briefly air dry and keep them at -20ºC. When I use them for staining, I air dry them, fix them in cold acetone for 5 minutes and start the staining process. I use the primary AB from BD Pharmingen, # 550285 at various dilution, incubate them overnight at 4ºC, use the secondary HRP conjugated antibody, DAB, counterstain and don't get any staining. Does anyone have any pointers, suggestions?

Thank you in advance,

Bea

Beatrice DeBrosse-Serra HT(ASCP)QIHC
Isis Pharmaceuticals
Antisense Drug Discovery
2855 Gazelle Ct.
Carlsbad, CA 92010
760-603-2371