TNFRI staining

Maria Denis maria_denis at yahoo.com
Wed Mar 24 03:03:55 EST 2004


Hi all,

I have a question regarding mouse macrophage staining for TNFRI(p55,
CD120a). We have tried the Pharmingen CD120a mAb in a 3-step labeling
protocol with a secondary biotinylated anti-hamster followed by
streptavidin-PE staining. The results we get are not reproducible,
and we have problems with high background staining, making difficult
the distinction between p55 wild type, heterozygote and knock out
cells. We are thinking of switching to a directly conjugated
anti-CD20a antibody instead, to avoid problems of non-specific
staining with the secondary antibody. 
Do you have any suggestions regarding what are the best antibodies
available for TNFRI staining and also what would be the best staining
protocol including blocking and FACS buffers?
Also, what other mouse cell types, besides macrophages, express
detectable levels of TNFRI?
Thank you in advance,

Maria 




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