Cd33 apc clone ac104.3e3

Flow cytometric analysis of hematopoietic surface markers was analyzed on a FACS Canto II (BD Biosciences, Franklin Lakes, New Jersey, USA) with the following antibodies: CD3-PerCP-Cy5.5 (clone UCHT1), CD19-FITC (clone HIB19), CD31-PE (clone WM59), CD34-APC (clone 581), CD38-PE (clone HIT2), CD43-FITC (clone 1G10), CD14-PE (clone MOP9), CD66b-PE (clone G10F5) (all BD Bioscience), CD45-APC-Cy7 (clone 2D1), CD117-PE-Cy7 (clone 104D2), CD11c-PE-Cy7 (clone 3.9), HLA-DR-FITC (clone LN3), CD235a-Pacific Blue (clone 6A7M) (all eBioscience, San Diego, California, USA), CD33-APC (clone AC104.3E3; Miltenyi Biotec). EBs were dissociated with Accutase (Stemcell Technologies, Vancouver, British Columbia, Canada) for 10–15 min prior to staining with antibodies. Single cells were incubated with 1% human IgG solution (Privigen, CSL Behring, King of Prussia, Pennsylvania, USA) for 30 min at 4 °C to block unspecific binding.
Immunophenotypic analysis of MSCs and iMSCs was performed with CD14-APC (clone M5E2), CD29-PE (clone MAR4), CD31-PE (clone WM59), CD34-APC (clone 581), CD45-APC (clone HI30), CD73-PE (clone AD2), CD90-APC (clone 5E10) (all BD Biosciences), and CD105-FITC (clone MEM-226; ImmunoTools, Friesoythe, Germany). Data was analyzed using the FlowJo software (Tree Star, Ashland, Oregon, USA).

CFSE was used to monitor the number of cell divisions (Walenda et al, 2010 (link)). The immunophenotype of HSPCs was examined at day 5 after infection by staining with CD13-PE (clone WM15), CD34-APC (clone 581), CD38-PE (clone HIT2), CD45-V500 (clone HI30), CD56-PE (clone B159; all by Becton Dickinson), CD33-APC (clone AC104.3E3), and CD133/2-PE (clone AC141; both Miltenyi Biotec) with or without CFSE staining. Cells were analyzed using a FACS Canto II (BD) with FACS Diva software (BD).