To isolate hematopoietic stem and progenitor cells (HSPCs) for in vitro differentiation or CUT&RUN experiments, bone marrow cells were harvested from femurs and tibias of 2 to 4 month-old Bcl11bRFP/YFP mice. CD117 MicroBeads (Miltenyi Biotec) were used to enrich HPSCs which were frozen in 90% FBS and 10% DMSO at 106 cells/mL. For CUT&RUN experiments, HPSCs were further purified by staining with anti-CD117 APC-eFluor780 (ThermoFisher Scientific) and with biotinylated antibodies against a panel of bone marrow lineage markers (CD19, CD11b, CD11c, NK.1.1, Ter119, CD3ε, Gr-1 and B220 (BioLegend)). Cells were then washed with HBH (Hank Balanced Salt Solution (HBSS) with 0.1% bovine serum albumin and 10mM HEPES) and stained with streptavidin-PerCP/Cy5.5 (BioLegend).
Anti cd117 apc efluor780
Manufactured by Thermo Fisher Scientific
Anti-CD117 APC-eFluor780 is a fluorescent-conjugated antibody that binds to the CD117 (c-Kit) cell surface antigen. CD117 is expressed on hematopoietic stem cells, mast cells, and other cell types. This product can be used in flow cytometry applications to detect and analyze CD117-positive cells.
Automatically generated - may contain errors
2 protocols using anti cd117 apc efluor780
1
Isolation of Hematopoietic Stem and Progenitor Cells
2
Isolation of Hematopoietic Stem and Progenitor Cells
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To isolate hematopoietic stem and progenitor cells (HSPCs) for in vitro differentiation or CUT&RUN experiments, bone marrow cells were harvested from femurs and tibias of 2 to 4 month-old Bcl11bRFP/YFP mice. CD117 MicroBeads (Miltenyi Biotec) were used to enrich HPSCs which were frozen in 90% FBS and 10% DMSO at 106 cells/mL. For CUT&RUN experiments, HPSCs were further purified by staining with anti-CD117 APC-eFluor780 (ThermoFisher Scientific) and with biotinylated antibodies against a panel of bone marrow lineage markers (CD19, CD11b, CD11c, NK.1.1, Ter119, CD3ε, Gr-1 and B220 (BioLegend)). Cells were then washed with HBH (Hank Balanced Salt Solution (HBSS) with 0.1% bovine serum albumin and 10mM HEPES) and stained with streptavidin-PerCP/Cy5.5 (BioLegend).
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