Dear Lori and group, We use Ig of the same species as the second step Ab. For example, if our first step Ab is rabbit, and the second Ab is goat anti-rabbit, we block first beffore staining with goat Ig from Sigma, at 10 mg/ml or sometimes higher concentrations. We have found that this treatment eliminates a lot of background, even for intracellular applications. We have used it in human, mouse, and rat cells, although we have not looked at whole blood, but either lines or cells from lymphoid organs. For single-step staining, use Ig of the same species as the Ab. Good luck. Beverly Barton, Ph.D. Dept. of Surgery UMD-New Jersey Medical School Newark, NJ 07103 USA Lori Krueger wrote: > > Dear Parisa, > > I am also interested in Fc receptor blockade and would appreciate hearing > what other investigators are using (besides Fab fragments and FcR specific > monoclonals). I predominantly work with platelets and have found that some > monoclonals augment platelet activation via the FcR. > Thanks, > > Lori Krueger > Center for Platelet Function Studies > UMass Medical School > > ----- Original Message ----- > From: "Amjadi, P" <p.amjadi@ic.ac.uk> > To: cyto-inbox > Sent: Friday, June 22, 2001 10:36 AM > Subject: FC block in immunophenotyping of human leukocytes > > > > > Dear All, > > We have observed quite a high background staining when we use fluorochrome > > conjucated mAbs for immunophenotyping of human machrophages and dendritic > > cells. I am looking for a FC block reagent in order to reduce the > > background staining. I checked the Pharmingen/BD catalogue and they only > > provide the reagent for rat and mouse and not for human. I would > appreciate > > if someone could help me with this query. > > > > Regards > > Parisa
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