Mouse bone marrow cells were enriched using a CD117 microbeads kit (Miltenyi Biotec) and separated using a MACS (Miltenyi Biotec) cell separator. After enrichment of CD117 cells, 2000 cells/well were seeded in Methocult (M3434) medium (STEMCELL Technologies). The methoCult medium-cell mixture was plated into 12-well plates with water supply in the inter-well chamber. After 7 days (1 passage) of incubation at 37C in 5% CO2, colonies were counted in 3 replicates per sample. The process was repeated for a total of 5 passages.
Cd117 microbeads kit
Manufactured by Miltenyi Biotec
The CD117 microbeads kit is a laboratory tool designed for the magnetic separation and purification of CD117-positive cells from a heterogeneous cell population. The kit contains superparamagnetic microbeads coated with anti-CD117 antibodies, which specifically bind to cells expressing the CD117 antigen. This allows for the efficient isolation and enrichment of CD117-positive cells for further analysis or downstream applications.
Automatically generated - may contain errors
Lab products found in correlation
Magnetic activated cell sorting (macs), by Miltenyi Biotec (1 mentions)
Methocult m3434 medium, by STEMCELL (1 mentions)
Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions)
Stem cell factor (scf), by Thermo Fisher Scientific (1 mentions)
M csf, by Thermo Fisher Scientific (1 mentions)
Macs lineage cell depletion kit, by Miltenyi Biotec (1 mentions)
2 protocols using cd117 microbeads kit
1
Mouse Bone Marrow Cell Culture
2
Lineage Marker Negative Cell Isolation and Differentiation
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Lineage marker negative cells (Lin−) were isolated from bone marrow or spleen using a MACS lineage cell depletion kit from Miltenyi Biotec. c-Kit+ and c-Kit− cells were separated by MACS using a CD117 MicroBeads kit from Miltenyi Biotec. Purity was assessed by labeling cells with the non-competing anti-c-Kit antibody (CD117; clone 3C11) from Miltenyi Biotec. Where indicated, purified cells were labelled with CFSE (Invitrogen). For obtaining HSPC-derived macrophages, purified Lin- cells were cultured in complete cell culture medium supplemented with 20 ng/ml Stem Cell Factor (SCF, Preprotech) and 50 ng/ml M-CSF (Preprotech) for 7 days.
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