Pe cy7 conjugated anti human cd4 mab

The expression of peptide-specific T cell receptors was examined on FACS-Canto II (Becton Dickinson, San Jose, CA) using PE-conjugated peptide/MHC tetramer (Medical and Biological Laboratories, Nagoya, Japan) according to the manufacturer's instructions. Briefly, in vitro expanded CTLs were incubated with peptide/MHC tetramer at room temperature for 10 min, and then a FITC-conjugated anti-human CD8 mAb, APC-conjugated anti-human CD3 mAb, PE-Cy7-conjugated anti-human CD4 mAb and 7-AAD (BD Biosciences) were added and incubated at 4°C for 20 min. HIV peptide (ILKEPVHGV)/HLA-A*02: 01 tetramer was used as a negative control.

Expression of peptide specific T cell receptors was analyzed on FACS-CantoII (Becton Dickinson, San Jose, CA) using URLC10, TTK, KOC1, VEGFR1 or VEGFR2-derived epitope peptide-MHC dextramer (Immudex, Copenhagen, Denmark) or tetramer-PE (Medical & Biological Laboratories Co.,Ltd., Nagoya, Japan) according to the manufacturer’s instructions. HIV-derived epitope peptide (RYLRDQQLL)-MHC dextramer or tetramer-PE was used as a negative control. T cells were incubated with peptide-MHC dextramer or tetramer-PE, treated with FITC-conjugated anti-human CD8 mAb, APC-conjugated anti-human CD3 mAb, PE-Cy7-conjugated anti-human CD4 mAb, and 7-AAD (BD Pharmingen, San Diego, CA.), and then analyzed with flow cytometry.