block for polyclonals

From:Linda Hylander

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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