I am trying to perform CFSE staining on fresh PBMC with CFSE (0.1uM). After culturing with a positive control peptide for 7 days I analyze via flow cytometry using PerCP to exclude CD14 CD19 and ViaProbe, CD4 on APC and I use a tetramer on PE.
I see a bright and dim CFSE population when viewing CFSE vs CD4 however tetramer is not showing up at all in CFSE vs PE.
Does anyone have experience with CFSE and tetramers? Could the CFSE be preventing my tetramer from binding to TCR?
Dear Alexandra Johansson
please check the attached methods paper, for tips & tricks that you may wanna follow.
Article A protocol for combining proliferation, tetramer staining an...
For a more detailed answer, could you share your FACS plots and protocol?
All the best,
Michael
Hi
I have done similar experiments before, CFSE doesn't interfere with PE tetramers for MHC-I. I assume you are using MHC-II tetramers for CD4 ! I think MHC-II tetramers are more challenging the MHC-I. Maybe you need to increase your CFSE concentration to see the proliferation.
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