A tetramer assay (tetramer stain) is used to detect and quantify antigen specific T cells. The interaction between a peptide-MHC complex (pMHC) and a cognate T cell receptor (TcR) is of too low affinity to be detected in flow cytometry. Streptavidin-mediated tetramerization of monomeric biotinylated pMHC (Altman JD. et al, 1996) increases the avidity of the pMHC streptavidin tetramer towards cognate TcRs and allow detection of antigen specific T cells.
For optimal results, please follow our MHC peptide loading protocol and tetramer staining protocols developed to obtain high pMHC complex yields with the easYmer® kits and subsequently stain T cells:
HLA easYmers® – tetramer production protocol
H2 easYmers® – tetramer production protocol
Assessment of HLA-complex folding – FACS assay
Protocol for staining human CD8+ T cells using peptide-HLA class I tetramers.
Protocol for staining murine CD8+ T cells using peptide-H2 class I tetramers.
Protocol for staining human CD4+ T cells using peptide-HLA class II tetramers.