Anti human cd49f fitc

Mammospheres from each passage of the breast normal tissues, primary tumors, and MCF-7 cell line were collected, washed, and stained according to the manufacturer's protocol. After generating a single-cell suspension, the detached cells were re-suspended in ice-cold PBS with 0.5% FBS at a density of 1 × 10 6 cells/200 μL. While the stem-like cells obtained from primary tumors and MCF-7 cell line were stained against human CD44-FITC, CD24-PE, the mammosphere-enriched normal cells were incubated with anti-human CD49f-FITC, and anti-human EpCAM-PE (all from BD Biosciences) at 4 °C in the dark for 45 min at recommended concentrations, according to the manufacturer's instructions. The labeled cells were then washed in PBS to remove any unbound antibodies, analyzed in FACS ARIA-2 (BD Biosciences, USA), and the data were visualized through DIVA software (BD Biosciences, USA). A minimum of 10,000 events was recorded for all samples and the experiments were performed in triplicate.