block for polyclonals
I was wondering
if anyone has suggestions to eliminate background when using
polyclonal Ab. I work with frozen cell lines and tissues, I rarely do
formalin fixed IHC.
Problem: I titered out a primary polyclonal Ab fine
with no background on a frozen cell pellet, acetone fixed slides, Ab
concentration 2.5ug/mL. I stained frozen tumor breast tissues, had loads of
background on my isotype control sections. My secondary Ab is goat anti-rabbit,
biotinylated, I blocked with 10% goat serum, avidin and BSA. I appreciate
any ideas and many thanks.
Linda
Hylander HT, IHC(ASCP)
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