Non-specific binding could be due to 1) dead cells taking up reagents (solution: gate out using propidium iodide), 2) way too much reagent (solution: titrate) 3) Fc receptors on your target cells (solution: use 24.G2 [FcBlock]), 4) CyChrome (Cy5PE) can stick (sometimes 24.G2 helps). Rachel M. Gerstein, Ph.D. Asst. Professor Dept. of Mol. Gen. & Micro. University Of Massachusetts Medical School 55 Lake Avenue North Worcester, MA 01655 508-856-5920 (FAX) Dr. Alwina Bittner wrote: > Hi collegues, > please accept me as a new member of the FACS-User-Group. > My first question: Has anyone experience in staining mouse splenocytes? I > have a problem with the high unspecific binding of several antibodies. Does > anybody know a method how to overcome this Problem. > > Thanks in advance > > Alwina > Dr. Alwina Bittner > MZ Nervenheilkunde - Neurologische Klinik mit Poliklinik > Neuroimmunologisches Labor > Rudolf-Bultmann-Str.8, 35039 Marburg, Germany, > Fax +49 6421 28 7055, Phone +49 6421 28 5480
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