RE: surface staining macrophages
P.Openshaw Medicine (p.openshaw@ic.ac.uk)
Wed, 26 Jul 1995 18:19:16 +0000 (GMT)
brett@borcim.wustl.edu (brett) wrote:
> 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?
We have stained lung macrophages using a mouse FcR block Pharmingen cat no
01241, which is good for anti-macrophage conjugated reagents. Must always
use an isotype control on each sample, which can be problematic. FITC label
of FAb may impare specific binding.
Do post other solutions.
*******************************************************************
*Peter Openshaw p.openshaw@ic.ac.uk *
*Respiratory Medicine Tel:+44 171 723 1252 x 5781/6 *
*St. Mary's Hosp. Med. School Fax:+44 171 724 7349 *
*Imperial College, Norfolk Place, London W2 1PG, UK *