Anti cd146pe clone 128 018

Characterization of ECFC was performed using speci c antibodies conjugated to uorochromes [ uorescein isothiocyanate (FITC), phycoerythrin (PE), peridin chlorophyll protein (PerCP)] that detect speci c endothelial surface markers: anti-CD31-FITC, clone MBC 78.2 (Invitrogen, Camarillo, CA, USA), anti-CD144-PE, clone TEA 1/31 (Beckman Coulter, Marseille, France), anti-CD146PE, clone 128018 (R&D Systems, Minneapolis, MN, USA), anti-VEGF R2/KDR-PE, clone 89106 (R&D Systems, Minneapolis, MN, USA), anti-CD34-FITC, My10 clone (BD, San Jose, CA, USA), anti-CD45-PerCP, clone 2D1 (BD, San Jose, CA, USA), and anti-CD133-APC, clone AC133 (Miltenyi Biotech, Auburn, CA, USA). The cytometric tubes were incubated for 30 min at 4 °C, protected from light, washed with PBS, centrifuged at 450 g for 5 min (RT), and then re-suspended in 300 µL of PBS for the acquisition of 10,000 events on a cytometer ow (FACS Calibur, Immuno uorometry Systems, Mountain View, CA, USA). Data analysis was performed using BD FACS DIVA 7.0 software (San Jose, CA, USA). Cells were considered to be ECFC when they were positive for CD31, CD144, CD146, and KDR markers, negative for CD45 and CD133, and showed decreased CD34 expression.