Anti ctla 4 clone bni3

Spleen or tumor tissues were mechanically disrupted and then filtered through a 70μM cell strainer (Fisher Scientific) to generate a single cell suspension. The cells were pelleted, washed in PBS, and then diluted into PBS at a concentration 2–4 million/ml. Staining was performed using fluorescently-labeled monoclonal antibodies specific for human cell surface markers, and compared to isotype-matched negative control antibodies used for fluorescence minus one (FMO) conditions. Antibodies used for staining were purchased from BioLegend, and included the following: anti-HLA-A,B,C (clone W6/32), anti-CD45 (clone HI30), anti-CD3 (clone OKT3), anti-CD19 (clone HIB19), anti-CD20 (clone 2H7), anti-PD-L1 (clone 29E.2A3), anti-PD-L2 (clone 24F.10C12), anti-PD-1 (clone EH12.2H7), and anti-CTLA-4 (clone BNI3 from BD Biosciences). Compensation was performed using single color controls, and staining was detected using an LSRII flow cytometer (BD Biosciences). Flow cytometric data analysis was performed using FlowJo software.