BioLegend
PE-Cy7 anti-mouse CD117 (c-Kit) Antibody
105813
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In order to track differentiation of myeloid cells from progenitors, I used CD117 (c-kit) to marker the cells in a progenitor status. CD117 is the receptor for stem cell factor (SCF), which is an important cytokine in myelopoiesis/hematopoiesis. In adult mice hematopoiesis occurs in the bone marrow; thus, I tracked CD117 expression on cells collected from the marrow. I titrated anti-murine CD117-PE-Cy7 on bone marrow leukocytes in the concentration range suggested by BioLegend. Most progenitor cells are lineage (Lin) negative, so I gated on cells that were CD90-, B220-, CD11b-, Ly6C-, Ly6G-. Nearly 20% of the Lin- cells were positive for CD117 (c-kit). While the population size did not change at the dilutions tested, the highest concentration (1:50) showed much greater background staining. The 1:200 dilution stained well with little background staining and is the concentration I have chosen to use on subsequent experiments.
Flow Cytometry
Murine bone marrow leukocytes
30 minutes at 4 C
Buffer contained BSA and FcBlock
Cells were fixed with 2% paraformaldehyde
None
Fluoresecence in flow cytometry
The gated Lin- cells, which includes the progenitor cells of the marrow, stained between 18.7 and 19.0% positive for CD117 for 1:50 to 1:200 dilutions. The 1:50 dilution had increased background staining of the negative population. The 1:200 dilution of anti-CD117 PE-Cy7 had the greatest proportion of CD117+ cells and appeared to have the least amount of background staining. The staining control without anti-CD117 showed no positive cells.
N/A
Works well at range of concentrations
Anti-CD117 PE-Cy7 is fine to use in your staining cocktails at 1:200 dilution.