Apc conjugated anti human cd133

The surface expression of CD133, CD44, and CD24 was analyzed by immunophenotyping. Briefly, 1X10⁶ cells were seeded in a 10 cm dish and incubated for 24 h in CO₂ incubator at 37°C prior to treatment. Cells were treated with TXL, APC, and TG alone or in combinations for 24 h. Cells were labeled with APC-conjugated anti-human CD133 (Miltenyi Biotec, San Diego, USA). FITC-conjugated mouse anti-human CD44 (BD Pharmingen, BD Biosciences, USA) and/or APC-conjugated CD24 in a buffer containing (1X PBS, 0.5% BSA and 2 mM EDTA) for 15 min at 4°C. Labeled cells were re-suspended in 1X PBS (pH 7.4), and analyzed by flow cytometer (LSR II A, BD Bioscience). Unstained cells served as negative controls. A total of 100,000 events were capture for each sample. Estimation of ALDH bright (ALDH^br) cell populations was performed 24 h after treatment using the Aldefluor^™ kit (Stem Cell Technology). Data were plotted using Win List 3D software.