Dear Amy, we usually study guinea pig and rat mast cell activation by measuring histamine release (fluorimetrically) and human basophil activation both through histamine release and CD63 or CD203c expression (thanks to the folks of this list who suggested it!). We also did several experiments by using CD203c in mast cells with some appreaciable results. Unfortunately the antibody (which works well with human basophils) does not seem to perfectly cross react with guinea pig or rat mast cells. Aniway it should be perfect with human mast cells. Good luck, Alfredo ______________________________________________ Dr Alfredo Vannacci, MD University of Florence Department of Preclinical and Clinical Pharmacology Viale Pieraccini,6 50139 Florence Italy > -----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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