Peptide binding assay by flow cytometry

HEK-293T cells were transiently transfected with pIRES2-EGFP-mClec9a plasmid, or the mock pIRES2-EGFP plasmid for binding assay. Cells (3 × 10⁵) were incubated with rat serum to block Fc receptors and different biotinylated peptides for 30 min on ice. After washing three times, cells were stained with streptavidin (SA)-PE for 30 min on ice, and the samples were analyzed for binding affinity by a flow cytometry after washing once.