Pe cy7 anti mouse cd133 antibody

ALDH activity was measured using an AldeRed ALDH detection assay kit (EMD Millipore SCR150) following the manufacturer’s instructions. Stained cells were analyzed using LSR II Green (BD Biosciences) with the green fluorescence channel (515–545 nm), and data were analyzed using the FACS DIVA software program (BD Biosciences). For sorting, single mouse primary acinar cells were incubated for 30 min with 100 μg/mL fluorescein isothiocyanate (FITC)-conjugated UEA-1 (Vector laboratories Cat# FL-1061–5) at room temperature in serum-free DMEM media (containing 0.1 mg/ml trypsin inhibitor and 10 μg/mL of DNase I). After washing, UEA1 FITC-labeled single acinar cells were stained with PE-Cy7 anti-mouse CD133 antibody (Biolegend 141209) in HBSS media (containing 10 mM HEPES, 5% FBS, 10 μg/mL of DNase I and 0.1 mg/ml trypsin inhibitor) for 20 min at 4°C. After washing, cells were stained with DAPI and analyzed/sorted using FACS Aria (BD Biosciences). In some experiments, acinar cells were stained with UEA1-FITC, AldeRed (Millipore, Cat# SCR150) or Super Bright 600 Anti-mouse cKit antibody (eBioscience, Cat# 63–1171-80), and PE-Cy7 anti-mouse CD133 antibody (Biolegend 141209).