Re: surface staining macrophages

/G=Gerhard/S=Nebe-von-Caron/OU=1890CHPE/O=TMGB.URC/@LANGATE.gb.sprint.com
Wed, 26 Jul 1995 08:06:00 -0400

If you know the isotype and subclass of your antibody, you
might also get away with an "unrelated antibody" block of a
different subclass and detect your reagent with a subclass
specific second step reagent (for example Pharmingen).



Gerhard Nebe-v.Caron
Unilever Research
Colworth Laboratory, UK

______________________________ Reply Separator _________________________________
Subject: surface staining macrophages
Author: brett@borcim.wustl.edu at INTERNET
Date: 25/07/95 19:54

Hello. I'd like to look at expression of a surface protein on mouse macrophage-
like cell lines by flow cytometry. I'm worried about decorating them with our
mouse mAb, which could be bound by FcR's on these cells. Two options I thought
of were 1) absorb mouse myeloma proteins to the FcR's, and FITC-label by mAb;
2) cleave the mAb into Fab or F(ab')2 fragments which could be FITC-conjugated.

What have other people done in these circumstances? What works and what doesn't
?

Thanks in advance,



Brett Lindenbach

Program in Immunology
Washington University - St Louis
brett@borcim.wustl.edu