Re: Fc Blocking
Simon Monard (Simon_Monard@adarc.org)
Wed, 18 Jun 1997 11:49:30 -0400
What I have always done is to pre-incubate your cells with 2% heat
inactivated human AB serum for 15 minutes at room temp. I suspend my cells
in PBS with the 2% serum plus a tad of azide, dispense into 12X75mm tubes,
spin them, remove all but about 20ul supernatent, resuspend the cells then
go for a cup of coffee. You then add your antibodies without washing. If
you are doing indirect staining use the above solution for your washing
before but not after your secondary antibody. There, easy. Sigma sell human
AB serum, heat inactivate it for 30 minutes at 56 degrees, aliquot and
freeze.
Simon Monard
ADARC
NYC
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