RE: Mast cells and basic orange 21

From: Wayne Shreffler (wayne.shreffler@mssm.edu)
Date: Tue Oct 29 2002 - 08:24:11 EST


We follow surface expression of CD63 and CD203c on basophils as an
indicator of activation.

I know that CD63 correlates well with mediator release (histamine) and I
have done a couple of experiments with 203c (after being pointed in the
right direction to find the Ab by members of this list) in which I've
seen strong upregulation in response to anti-IgE consistent with the
literature.

I know that there is literature on variable expression of CD63 on mast
cells of different origin or in mastocytosis and I think people have
shown that it is upregulated upon degranulation, but I can't swear to
it.

CD203c is expressed only on basophils and mast cells according to the
literature. It is upregulated on basophils upon activation. You might
try it for your mast cells line.

Wayne

----------------------------------------------------------
Wayne G. Shreffler, MD, PhD
Fellow, Allergy & Clinical Immunology
Mount Sinai Medical Center

-----Original Message-----
From: Amy Raber [mailto:araber@athersys.com]
Sent: Friday, October 25, 2002 2:58 PM
To: cyto-inbox
Subject: Mast cells and basic orange 21


Dear Flow users,

I am interested in stimulating a mast cell line and using flow cytometry
to screen for degranulation.  I came across mention of a dye-basic
orange 21 that may be appropriate for this assay.  I was hoping someone
would know where I can purchase this dye?  Is there a protocal somewhere
for staining?  Or is there a better method to measure degranulation?
(we are looking into annexin staining for degranulation too)  We only
have access to 488nm laser.


Thanks so much for any help you can send my way!!

Amy

Amy Raber
Athersys Inc.
Cleveland, OH



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