Fitc conjugated mouse anti human primary pd l1 monoclonal antibody

A 0.5–1 × 10⁶ cultured cells were collected using cell scrapers and resuspended in 100 μL incubation buffer (0.25 g bovine serum albumin dissolved in 50 mL 1 × PBS) with 20 μL of FITC-conjugated mouse anti-human primary PD-L1 monoclonal antibody (BD Pharmingen, P/N 558065) for 1 h at room temperature. Cells were rinsed 2× in incubation buffer and finally resuspended in 0.45 mL ice cold 1× PBS and strained into polystyrene flow cytometry tubes. Flow Cytometry was conducted using the BD FACScan Flow Cytometer and each experiment was conducted in triplicate. Examples for gating parameters can be found in Supplementary Figure 1. Relative fluorescence intensity (RFI) was calculated by normalizing mean fluorescence intensity (MFI) measurements of treated samples to vehicle control:

0.5–1 × 10⁶ cultured cells were collected with cell scrapers and resuspended in 100 µL incubation buffer (0.25 g bovine serum albumin in 50 mL 1 × PBS) with 20 µL of FITC-conjugated mouse anti-human primary PD-L1 monoclonal antibody (BD Pharmingen, P/N 558065) for 1 h at room temperature. Cells were rinsed twice in incubation buffer and finally resuspended in 0.45 mL ice cold 1 × PBS and strained into polystyrene flow cytometry tubes. Flow Cytometry was conducted using the CytoFLEX S Flow Cytometer–Beckman Coulter and each experiment was conducted in triplicate.