Protocols
To ensure efficient and specific staining with our peptide-MHC reagents, we have developed several dedicated staining protocols for CD4+ and CD8+ T cells.
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:
easYmers®
easYmer® – tetramer production protocol
Assessment of HLA-complex folding – FACS assay
Use this Support spreadsheet to calculate volume to prepare different test amounts
CD8+ T cells Tetramer staining protocol
Protocol for staining CD8+ T cells using peptide-MHC class I tetramers.
CD4+ T cells Tetramer staining protocol
Protocol for staining CD4+ T cells using peptide-MHC class II tetramers.