FC blocking is it really necessary?

From: Derek Schulze (flow@post.queensu.ca)
Date: Fri Jan 28 2000 - 18:26:50 EST


We are not seeing a difference between FC gamma RII/III blocked and
unblocked samples for flow analysis of mouse bone marrow or mouse
splenocytes.  We are also not seeing high background in our histograms.

For example, when looking at CD4 in splenocytes we see a tightly grouped
negative population in the first decade on the log scale with an even
tighter CD4 (36%) positive population in the third decade.  No broad spread
between them to speak of.  In the Pharmingen catalogue they show an assay
for their Fcgamma-block that shows a broad peak showing 100% staining for
Thy1.2 in the absence of blocking.  Any ideas why we see no effect?

We have achieved similar results with Ly-6G x CD11b, Ter119 x CD44 and B220
x IgM - absolutely no difference between FC blocked or not blocked.

Why do we keep seeing references to blocking FC receptors in the literature
when we don't seem to be having a problem with background to begin with?
Are we doing something wrong?

- Derek Schulze

Flow Cytometry and Confocal Microscopy Core Facility
Cancer Research Labs
Queen's University
Kingston, Ontario
Canada



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