Quantitating MHC class I ligand production and presentation using TCR-like antibodies.

摘要: Accurately determining the number of peptide-MHC class I complexes on cell surface is necessary when evaluating cellular processes or pharmaceuticals that alter antigen presentation machinery. Here describe a quantitative flow cytometry application for cells grown in tissue culture express an endogenous protein from which peptide derived. The procedure requires monoclonal antibody with ability to distinguish MHC molecules presenting interest other complexes. Fluorescence signal measured antibody-labeled can be compared fluorescent-calibrated beads determine relative antibodies bound and hence specific expressed by cell. As new TCR-like specificity are created, this method will helpful quantifying exact numbers generated types relating these physiological outcomes T activation.