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RB780 Mouse Anti-Human CD117 (c-Kit)
RB780 Mouse Anti-Human CD117 (c-Kit)
Flow cytometric analysis of CD117 (c-Kit) expression on Human TF-1 Cells. Cells from the Human TF-1 (Human erythroleukemia, ATCC® CRL-2003™) cell line were stained with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; dashed line histogram) or BD Horizon™ RB780 Mouse Anti-Human CD117 (c-Kit) antibody (Cat. No. 569063/569064; solid line histogram). DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) 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 (DAPI-negative) TF-1 cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Flow cytometric analysis of CD117 (c-Kit) expression on Human TF-1 Cells. Cells from the Human TF-1 (Human erythroleukemia, ATCC® CRL-2003™) cell line were stained with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; dashed line histogram) or BD Horizon™ RB780 Mouse Anti-Human CD117 (c-Kit) antibody (Cat. No. 569063/569064; solid line histogram). DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) 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 (DAPI-negative) TF-1 cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Product Details
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BD Horizon™
KIT; c-Kit; SCFR; PBT; Mast/stem cell growth factor receptor
Human (QC Testing)
Mouse BALB/c IgG1
Megakaryoctic cell line MOLM-1
Flow cytometry (Routinely Tested)
5 µl/test
VI C30
3815
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed.

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.

Product Notices

  1. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. 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.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
  11. For U.S. patents that may apply, see bd.com/patents.
569064 Rev. 2
Antibody Details
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104D2

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.

569064 Rev. 2
Format Details
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RB780
The BD Horizon RealBlue™ 780 (RB780) Dye is part of the BD family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 781-nm. Driven by BD innovation, RB780 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.
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RB780
Blue 488 nm
498 nm
781 nm
569064 Rev.2
Citations & References
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View product citations for antibody "569064" on CiteAb

Development References (8)

  1. 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). View Reference
  2. 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.
  3. 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
  4. 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
  5. Kinashi T, Springer TA. Steel factor and c-kit regulate cell-matrix adhesion. Blood. 1994; 83:1033-1038. (Biology). View Reference
  6. 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: Flow cytometry). View Reference
  7. 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
  8. 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
View All (8) View Less
569064 Rev. 2

 

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.