All MHC class I easYmer® molecules are biotinylated allowing you to make tetramers with your preferred labels. Stored frozen, MHC class I easYmers® are stable and peptide receptive for 24 months; once complexed with peptides, the resulting monomers are stable for years in the freezer.
We recommend that you use predictors (e.g. NetMHC series or NetMHCpan) as an aid to design your monomers. If the specific peptide-MHC class I combination of interest is not predicted to bind with a sufficiently low %Rank, there’s a high risk that the peptide will not support the formation of stable complexes. Thus, to ensure proper peptide loading and optimal staining of your CD8+ T cells, we strongly recommend that you experimentally verify successful peptide-MHC (pMHC) complex formation. To this end, the easYmer kit includes a simple FACS based protocol allowing you to validate proper peptide loading (click here for protocol).
The peptide loaded MHC monomers can be long-term stored frozen for later tetramerisation. When needed, these monomers can easily be tetramerised with labelled streptavidin (e.g. fluorophore label for analysis in a classical flow cytometry assay or metal-labels for analysis by CyTOF). For optimal HLA tetramer staining of CD8+ T cells, please follow our MHC class I tetramer staining protocol.
Add peptide to easYmers® and incubate. Add streptavidin-conjugate and stain T cells.