Pe cy7 conjugated anti human cd34

Transduction with the AdV5/35-ZFNs and the donor IDLV, was analyzed by flow cytometry analysis (FACSCalibur; Becton Dickinson Pharmingen). Immunophenotypic analysis of the hematopoietic differentiated cells was performed using the following antibodies according to the manufacturer's instructions: phycoerythrin (PE)-Cy7-conjugated anti-human CD34 (BD Pharmingen), PE-conjugated anti-human CD31 (eBiosciences), allophycocyanin (APC)-conjugated anti-human CD45 (BD), and fluorescein isothiocyanate (FITC)-conjugated anti-human CD43 (BD). Fluorochrome-matched isotypes were used as controls. 4′,6-Diamidino-2-phenylindole (DAPI; Roche)-positive cells were excluded from the analysis. Analysis was performed using FlowJo software.

Human cells in the BM and PB of recipient mice were stained with Lineage mixture (biotinylated anti-human CD14, CD3, CD56, CD19 and CD235a), PE-Cy7-conjugated anti-human CD34, APC-conjugated anti-human CD45, PE-conjugated anti-human CD38, biotinylated anti-human CD7, biotinylated anti-human CD10, PE-conjugated anti-human CD62L, APC-Cy7-conjugated anti-human CD3, PE-conjugated anti-human CD11b, or PE-Cy7-conjugated anti-human CD19 (BD Biosciences, San Jose, CA, USA). They were then analyzed on a FACS Canto II or sorted on a FACS Aria II (BD Biosciences).