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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