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PE Mouse Anti-Human CD117 (c-Kit)
PE Mouse Anti-Human CD117 (c-Kit)

Flow cytometric analysis of CD117 expression on human TF-1 Cells. Cells from the human TF-1 (Human erythroleukemia, ATCC Cat. No. CRL-2003) cell line were stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dashed line histogram) or PE Mouse Anti-Human CD117 antibody (Cat. No. 567129/567132; solid line histogram). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histogram showing CD117 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) TF-1 cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.

Flow cytometric analysis of CD117 expression on human TF-1 Cells. Cells from the human TF-1 (Human erythroleukemia, ATCC Cat. No. CRL-2003) cell line were stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dashed line histogram) or PE Mouse Anti-Human CD117 antibody (Cat. No. 567129/567132; solid line histogram). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histogram showing CD117 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) TF-1 cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.

Product Details
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BD Pharmingen™
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
VI C30
3815
Aqueous buffered solution containing BSA and ≤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 unconjugated antibody and free dye were removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to 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 cell and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
567129 Rev. 1
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.

        

567129 Rev. 1
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
567129 Rev.1
Citations & References
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Development References (3)

  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. 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
567129 Rev. 1

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

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