Brief Protocol: |
After blocking with anti-CD16/32, microbeads against c-kit were used to enrich c-kit positive cells. After washing, cells were stained with many Alexa Fluor® 488 conjugated antibodies against mouse lineage markers (Ly-6G/Ly-6C (Gr-1)Cat# 108417, CD11b Cat# 101217, TER-119/Erythroid cells Cat# 116215,CD4 Cat# 100529, CD8a Cat# 100723, CD45R/B220 Cat# 103225, CD3e Cat# 100321, CD11c Cat#117311). c-kit, Sca-1, CD150, CD48 and CD135 were also stained for 15 min at 4ËšC. Cells were washed twice with staining buffer and then sorted for long term hematopoietic stem cell culturing. |
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