Re: Help to interplate some intriguing flow cytometry data

From: PeterG (gogy@freemail.hu)
Date: Fri Apr 19 2002 - 03:19:10 EST


> Xinjian Chen M.D., Ph.D." <xchen2@emory.edu> - 4/11/02
9:00 PM
>
> Greetings to everybody
> I have recently obtained some intriguing flow data, as
described below, and
> am looking for help to understand its nature. When I
perform multi-color
> staining on spleen or lymph node cells of immunized mice
with anti-CD4-FITC
> plus other antibodies, I can unequivocally identify by
flow cytometry a
> subset (fifteen to 30%) of CD4+ T cells that are positive
for B cell
> markers, including B200, IgM, IgD, MHC Class II, etc.
However, if I use same
> anti-CD4 antibody conjugated to other flurochrome, such
as PE or PerCP, than
> FITC for multi-color staining, the B cell marker-positive
subset of CD4 T
> cells becomes no longer identifiable.
> Your comments will be greatly appreciated.
>
>
> Best regards
> Xinjian

Hello Xinjian,

One possibility, the anti-CD4-FITC is contaminated with
some
"anti-mouse B marker-FITC" (eg. aCD19-FITC).
An unchanged pipette tip could be the reason.
Try a new batch of anti-CD4-FITC.

Best regards,

Gogy

Peter Gogolak, Ph.D.
Institute of Immunology
University of Debrecen, Hungary
Phone/Fax: +36 52 417159



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