Re: Latex beads for flow

From: adwells@mail.MED.UPENN.EDU
Date: Thu May 13 1999 - 12:51:02 EST


>Has anyone ever used protein attached to latex beads as a target for
>antibody evaluation by flow? I have a 19 KDa recombinant protein for
>Plasmodium falciparum that I want to use as a probe to look for IgG
>subclasses in human sera. Any suggestion (comercial source, protocol for
>protein attachement, any trick?????)

Olindo,

I have used 5 micron sulfated latex beads from Interfacial Dynamics Corp.
(product number 1-5000; ph 800-323-4810) as artificial APCs by coating them
with antibodies [proteins].  You just add your protein (in PBS @ ~10µg/ml)
to the beads (~10^7) and incubate for ~2 hours and block.  Such beads when
coated w/ protein can be used as targets for antibodies and their light
scatter and fluorescence properties can easily be assessed by flow
cytometry.

AW




Andrew D. Wells, Ph.D.
University of Pennsylvania
Department of Medicine
904 Stellar-Chance Laboratories
422 Curie Boulevard
Philadelphia, PA  19104
(215) 898-1951
(215) 573-2880 (FAX)
adwells@mail.med.upenn.edu



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