Anti human cd29 pe

Sub-cultured AMCs were trypsinized into a single cell suspension and maintained in serum-containing media at 37^oC for 60 min to allow the cells time to regenerate the surface markers damaged or lost during the trypsinization. Non-specific epitopes on cells were blocked using 4% NDS in phosphate-buffered saline for 30 min at room temperature followed by labeling cells in suspension with fluorescent-conjugated primary antibodies at 4^oC for 60 min in the dark. Primary antibodies were purchased from BD Biosciences and used at the concentrations indicated; anti-human CD29-PE (1:100), CD34-FITC (1.5:100), CD45-FITC (2.5:100), CD90-PE (1:200), CD31-FITC (1.5:100), CD44-PE (1:100), CD73-FITC (1:100) and CD105-PE (1:100) with their respective isotype controls (2.5:100). Cells were imaged on a fluorescence-activated cell sorter Calibur (Becton Dickinson, Franklin Lakes, NJ, USA). Gating was implemented after profiling unstained and isotype control cell samples.