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Flow cytometric analysis of CD117 (c-Kit) expression on Human TF-1 cells. TF-1 cells (Human erythroleukemia cell line; ATCC Cat. No. CRL-2003) were stained with either BD Horizon™ BUV395 Mouse IgG1, k Isotype Control (Cat. No. 563547; dashed line histogram) or BD Horizon™ BUV395 Mouse Anti-Human CD117 (c-Kit) antibody (Cat. No. 568489; solid line histogram) at 0.25 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histogram showing CD117 (c-Kit) expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ BUV395 Mouse Anti-Human CD117 (c-Kit)
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
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- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
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- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
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The 104D2 monoclonal antibody specifically binds to human CD117, the receptor for stem cell factor (SCF). It selectively recognizes NIH- 3T3 cells transfected with human c-kit, the gene that codes for SCF-R. The 104D2 antibody does not block the epitope that binds SCF. In the bone marrow of humans and mice, SCF is expressed primarily on hematopoietic progenitor cells. Lack of functional SCF or deficient SCF-R caused by mutations in the Sl and W loci, respectively, can result in severe anemia and a decrease in the number of primitive progenitor cells in mice. Human hematopoietic progenitor cells can be recognized by their surface expression of CD34. This cell population constitutes a small subset (1% to 5%) of bone marrow cells. CD34+ cells contain a small subpopulation of primitive/non-committed progenitors, with the remaining fraction being cells committed to the various hematopoietic lineages. SCF alone induces extensive proliferation of erythroid-committed progenitor cells (CD34lo CD71hi CD64-). On primitive (CD34hi CD38lo CD50+) and granulo-monocytic (CD34+ CD64+) progenitor cells, SCF synergistically enhances the effects of other cytokines, the strongest of which are on the primitive progenitor cells. In addition, SCF promotes survival of primitive progenitors in the absence of proliferation. The receptor is highly expressed at similar levels on all of the three mentioned CD34+ cell subsets, whereas B-lymphoid committed progenitor cells (CD34+ CD19+) express low levels of SCF-R. Among CD34- bone marrow cells, only a small number of cells (mostly erythroid) express the receptor.
Development References (8)
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Ashman LK, Buhring HJ, Aylett GW, Broudy VC, Muller C. Epitope mapping and functional studies with three monoclonal antibodies to the c-kit receptor tyrosine kinase, YB5.B8, 17F11, and SR-1. J Cell Physiol. 1994; 158(3):545-554. (Biology: Flow cytometry). View Reference
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Ashman LK, Cambareri A, Nguyen L, Bühring H-J. CD117 workshop panel report. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:816-818.
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Ikuta K, Weissman IL. Evidence that hematopoietic stem cells express mouse c-kit but do not depend on steel factor for their generation. Proc Natl Acad Sci U S A. 1992; 89(4):1502-1506. (Biology). View Reference
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Keller JR, Ortiz M, Ruscetti FW. Steel factor (c-kit ligand) promotes the survival of hematopoietic stem/progenitor cells in the absence of cell division. Blood. 1995; 86:1757-1764. (Biology). View Reference
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Kinashi T, Springer TA. Steel factor and c-kit regulate cell-matrix adhesion. Blood. 1994; 83:1033-1038. (Biology). View Reference
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Rappold I, Ziegler BL, Kohler I, et al. Functional and phenotypic characterization of cord blood and bone marrow subsets expressing FLT3 (CD135) receptor tyrosine kinase. Blood. 1997; 90(1):111-125. (Immunogen). View Reference
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Simmons PJ, Aylett GW, Niutta S, To LB, Juttner CA, Ashman LK. c-kit is expressed by primitive human hematopoietic cells that give rise to colony-forming cells in stroma-dependent or cytokine-supplemented culture. Exp Hematol. 1994; 22:157-165. (Biology). View Reference
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Yarden Y, Kuang WJ, Yang-Feng T, et al. Human proto-oncogene c-kit: a new cell surface receptor tyrosine kinase for an unidentified ligand. EMBO J. 1987; 6(11):3341-3351. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.