|From:||Linda Hylander |
----- Original Message -----From: rueggpTo: Linda HylanderSent: Saturday, February 23, 2002 11:29 AMSubject: Re: block for polyclonalsLinda,
John's reply reminded me that BSA can surely be your problem. I stopped using BSA and do much better with serum block both from the host of the secondary and serum from the species being stained. This would be like having a human absorbed secondary. I use this serum block before the primary antibody and then again before the secondary.
Linda Hylander wrote: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)