Apc labeled anti mouse cd45r b220

Murine B cells cultured at a concentration of 5 × 10⁵ cells/ml in RPMI medium supplemented with 15% FBS, penicillin-streptomycin (100 units/ml) and L-glutamine (2 mM) were treated with anti-CD40 antibody (1 μg/ml, eBioscience) and recombinant mouse IL-4 (20 ng/ml; PeproTech). Cells were collected at the indicated time points. Class switch recombination (CSR) was measured by staining with PE-labeled anti-mouse IgG1 (BD Biosciences) and APC-labeled anti-mouse CD45R/B220 (BD Biosciences). The expression of CD40 and FAS was measured by staining with APC-labeled anti-mouse CD40 (BioLegend) and APC-labeled anti-mouse CD95 (BioLegend), respectively. Data acquisition was performed using a FACSVerse flow cytometer (BD Biosciences) (Cheong et al., 2016 (link)).

Single-cell suspensions from normal and malignant spleen samples were fixed in ethanol and stained with APC-labeled Anti-Mouse CD45R/B220 (BD Pharmingen) and FITC labeled Anti-Mouse CD19 (BD Pharmingen) or PE labeled anti-mouse IgM (BD Pharmingen) to determine the abundant cell type present. Data acquisition was performed at the Flow Cytometry and Cellular Imaging Core Facility at MD Anderson on a FACSCalibur flow cytometer and analysis was performed with FlowJo Ssoftware.