Human cd44 apc

For immunofluorescence staining, 1 × 10⁶ cells were stained with APC/FITC-conjugated antibodies for 40 min at RT, as previously described [41 (link)]. Immuno-stained cells were then washed twice with PBS and resuspended in 500 μl of phosphate-buffered saline and used for flow analysis. The following primary antibodies were used: human CD235-APC (Cat.551336), human CD34-APC (Cat.555824), human CD69-APC (Cat.555533), human CD44-APC (Cat.559942), human CD71-FITC (Cat.555536), human CD235a-APC (Cat.551336) were all purchased from BD Biosciences (BD Biosciences, NJ, US). c-KIT-APC (17-1171-82) was purchased from eBioscience (eBioscience Inc, CA, US). Flow cytometry was performed using a NovoCyte flow cytometer (ACEC Biosciences Inc, CA, US) and Novo-express software.

Immunophenotyping assays were all analyzed on LSRII Flow Cytometer (Franklin Lakes, NJ, USA). Flow data were further analyzed by FlowJo software (TreeStar, Ashland, OR, USA). Antibodies were as follows: human CD44-APC and human CD45-PE (BD Pharmingen™, NJ, USA). Leukemic cells carrying green fluorescence proteins (GFPs) or red fluorescence proteins were detected and selected by MoFlo flow-sorter (Beckman coulter, Fullerton, CA, USA).