> -----Original Message----- > From: Antony Bakke [mailto:bakkea@ohsu.edu] > Sent: Friday, April 19, 2002 7:10 PM > To: Cytometry Mailing List > Subject: tetramer staining > > Our tetramer staining is giving significant background > staining in people who are > negative for the HLA-A2 allele and negative for exposure. > The background is low > intensity. Any suggestions about blocking this background staining? > You will probably need to work this out in your system, but here are some general ideas from my experience with tetramers: - titrate the tetramer - try preincubating the cells with FCS [we block for 5 mins with 2% FCS] - try labeling at different temperatures [we found 37oC superiour for reducing background whild still providing good labeling] - try whole blood labeling Some tetramers apparently have some specifici interaction with CD8, you may want to investigate if thsi is what you're seeing. Good luck, Tom **************************************************************************** * Thomas W. Mc Closkey, Ph. D. Director of Flow Cytometry, North Shore University Hospital Assistant Professor of Pediatrics, New York University School of Medicine Boas Marks Biomedical Research Center, 350 Community Drive Manhasset, Long Island, New York 11030 ph: 516-562-4844 [office], 516-562-1135/4641 [lab] fax: 516-562-2866 **************************************************************************** *
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