For isolation of BM LP cells, bones were crushed in a mortar, and the cell suspension passed through a 50-µm filter. For progenitor cell isolation by FACS sorting, BM cells were either sorted unenriched, enriched for KIT+ cells by magnet-activated cell sorting column for using anti-CD117 immunomagnetic beads (Miltenyi Biotec), or lineage depleted by using sheep anti–rat IgG beads (Dynabeads; Invitrogen), recognizing purified lineage antibodies against CD4 (GK1.5; BioLegend), CD8A (53–6.7; BioLegend), CD11B (M1/70; BioLegend), Ter119 (TER-119; BD Biosciences), GR1 (RB6-8C5; BioLegend), and CD19 (6D5; BioLegend) before antibody staining. Cells were thereafter stained with antibodies (full information of antibody combinations, clones, and conjugates is shown in Table S4). Cell sorting was performed on BD FACSAriaII or FACSAriaIII cell sorters (BD Biosciences) and analyses on a BD FACSLSRII, FACSAriaIII, FACSCantoII, or FACSFortessa.
Anti cd117 immunomagnetic beads
Manufactured by Miltenyi Biotec
Anti-CD117 immunomagnetic beads are a lab equipment product designed for the isolation and purification of CD117-positive cells. These beads are composed of a magnetic core coated with antibodies specific to the CD117 surface antigen. They can be used to selectively bind and separate CD117-expressing cells from complex samples, such as bone marrow or peripheral blood, for further analysis or downstream applications.
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Lab products found in correlation
Cd11b m1 70, by BioLegend (1 mentions)
Cd19 6d5, by BioLegend (1 mentions)
Gr 1 rb6 8c5, by BioLegend (1 mentions)
Cd4 gk1, by BioLegend (1 mentions)
Cd8a 53 6, by BioLegend (1 mentions)
Facs lsr 2, by BD (1 mentions)
Dynabead, by Thermo Fisher Scientific (1 mentions)
Facscanto 2, by BD (1 mentions)
Facsaria 3, by BD (1 mentions)
Facsaria 2, by BD (1 mentions)
Streptavidin qd655, by Thermo Fisher Scientific (1 mentions)
Cd16 32 93, by Thermo Fisher Scientific (1 mentions)
2 protocols using anti cd117 immunomagnetic beads
1
Isolation and Sorting of Bone Marrow Progenitor Cells
2
Cell Surface Marker Staining and Purification
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Antibodies used for cell surface staining were as follows: CD11b/Mac1 (M1/70), CD4 (H129.19), CD8a (53–6.7), B220/CD45 (RA3–6B2), CD5 (Ly1), Ter119 (Ter-1119), Gr1/Ly6G and Ly6C (RB6–8C5), CD19 (ID3), CD41/Itga2b (MWReg30), and CD135/Flt3 (A2F10.1) (104) (BD Biosciences Pharmingen) and NK1.1 (PK136), Sca1 (D7), CD117/c-Kit (2B8), CD16/32 (93), and CD105/Eng (MJ7/18) (eBioscience). Biotinylated antibodies were visualized with streptavidin-QD655 (Invitrogen) or streptavidin-tricolor (Invitrogen), and purified lineage antibodies were visualized with polyclonal goat anti-rat tricolor (Invitrogen) or polyclonal goat anti-rat-QD605 (Invitrogen). MACS column enrichment of c-Kit+ cells was done using anti-CD117 immunomagnetic beads (Miltenyi Biotec) as previously described [20 (link)].
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