Pe cd38 clone hit2

Human AML cells at a concentration of 1 million cells/mL were sorted on BD FACS Aria III to isolate immunophenotypically defined HSC (CD45^dimSSc^lowLin^–CD90⁺CD34⁺CD38^–), LSC-enriched (CD45^dimSSc^low Lin^–CD90^–CD34⁺CD38^–), and non-LSC-enriched (CD45^dimSSc^lowLin^–CD90^–CD34⁺CD38⁺) populations. The anti-human antibodies used for FACS analysis were FITC-CD34 (clone 561, Biolegend), PE-CD38 (clone HIT2, Biolegend), APC-CD45 (clone HI30, Biolegend), Pacific Blue-Lineage cocktail (#348805, Biolegend) combined with Pacific blue-CD138 (clone MI15, Biolegend) to exclude plasma cells, APC/Cy7-CD90 (clone 5E10, Biolegend), BV421-CD99 (clone 3B2/TA8, Biolegend), Pacific Blue-CD123 (clone 6H6, Biolegend), and APC-TIM3 (clone F38–2E2, Biolegend). Briefly, cryopreserved or fresh mononuclear cells from AML patients were stained with antibody cocktail on ice for 30 minutes before being washed once and resuspended in sorting buffer. A high purity single cell sorting strategy was used for cell numbers less than 500 into PCR strip tubes. Bulk leukemic blasts (>20,000 cells) were sorted into 1.5 mL Eppendorf tubes. Purity of sorting achieved for all samples were >98%.