Re: new member - unspecific antibody binding to mouse splenocytes

From: Rachel M. Gerstein (Rachel.Gerstein@ummc.ummed.edu)
Date: Fri Jan 08 1999 - 12:55:52 EST


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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