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PE-Cy™7 Rat anti-Mouse CD117 2B8 RUO

PE-Cy™7 Rat anti-Mouse CD117 

Clone  2B8   (RUO)

  • Brand BD Pharmingen™
  • Alternative Name c-KIT; W; SCFR; Stem Cell Factor Receptor; Sl; Steel Factor Receptor; Ssm
  • Concentration 0.2 mg/ml
  • Isotype Rat WI, also known as Wistar (outbred) IgG2b, κ
  • Reactivity Mouse (QC Testing) 
  • Application Flow cytometry (Routinely Tested) 
  • Immunogen Mouse Bone Marrow Mast Cells
  • Entrez Gene ID 16590 
  • Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
  • Regulatory Status RUO
Product Details Recommended Assay References

Description

The 2B8 monoclonal antibody specifically binds to CD117 (c-Kit), a transmembrane tyrosine-kinase receptor that is encoded by the Kit gene (formerly dominant white spotting, W). The c-Kit ligand (also known as steel factor, stem cell factor, and mast cell growth factor) encoded by the Kit1 gene (formerly steel, SI), is a co-mitogen for hematopoietic stem cells, myeloerythroid progenitors and a mast-cell differentiation factor. The KitW and Kit1SI mutant alleles have similar pleiotropic effects on the development of melanocytes, germ cells, and the hematopoietic system. In the adult bone marrow, CD117 is expressed on hematopoietic progenitor cells, including CD90 (Thy-1) low, TER-119-, CD45R/B220-, CD11b (Mac-1)-, Ly-6G (Gr-1)-, CD4-, CD8-, and Sca-1 (Ly-6A/E)+ multipotent hemotopoietic stem cells, progenitors committed to myeliod and/or erythroid lineages, and precursors of B and T lymphocytes. This widespread expression of CD117 in hematopoietic precursors is consistent with the participation of c-Kit and its ligand in the regulation of several hematopoietic lineages. Intrathymic expression of c-Kit and c-Kit ligand suggest that CD117 is also involved in the regulation of some events during the development of T lymphocytes. CD117 is also expressed by mast cells and by dendritic cells found in the periarteriolar lymphocytoc sheaths (T-cell areas) of splenic white pulp. The mAb 2B8 reportedly does not block the action of c-Kit. This clone 2B8 had been reported to cross-react with rat.

Format
  • Format PE-Cy™7
  • Excitation Source Blue 488 nm,Green 532 nm,Yellow/Green 561 nm
  • Excitation Max 496 nm
  • Emission Max 785 nm

PE-Cy™7 is a tandem fluorochrome that combines PE and a cyanine dye. PE-Cy7 conjugated reagents are as bright as PE conjugates. PE-Cy7 is particularly sensitive to photo-induced degradation, resulting in loss of fluorescence and changes in fluorescence spillover. Extreme caution must be taken to avoid light exposure and prolonged exposure to paraformaldehyde fixative. Fixed cells should be analyzed within 4 hours of fixation in paraformaldehyde or transferred to a paraformaldehyde-free buffer for overnight storage.

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Preparation and Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with PE-Cy7 under optimum conditions, and unconjugated antibody and free PE-Cy7 were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  5. Cy is a trademark of Amersham Biosciences Limited. This conjugated product is sold under license to the following patents: US Patent Nos. 5,486,616; 5,569,587; 5,569,766; 5,627,027.
  6. This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
  7. 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.
  8. Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
  9. 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.


  1. Anderson DM, Lyman SD, Baird A, et al. Molecular cloning of mast cell growth factor, a hematopoietin that is active in both membrane bound and soluble forms. Cell. 1990; 63(1):235-243. View reference

  2. Austen KF, Boyce JA. Mast cell lineage development and phenotypic regulation. Leuk Res. 2001; 25(7):511-518. View reference

  3. Domen J, Weissman IL. Hematopoietic stem cells need two signals to prevent apoptosis; BCL-2 can provide one of these, Kitl/c-Kit signaling the other. J Exp Med. 2000; 192(12):1707-1718. View reference

  4. Fadini GP, Sartore S, Schiavon M, et al. Diabetes impairs progenitor cell mobilisation after hindlimb ischaemia-reperfusion injury in rats. Diabetologia. 2006; 49(12):3075-3084. View reference

  5. Godfrey DI, Zlotnik A. Control points in early T-cell development. Immunol Today. 1993; 14(11):547-553. View reference

  6. Huang E, Nocka K, Beier DR, et al. The hematopoietic growth factor KL is encoded by the Sl locus and is the ligand of the c-kit receptor, the gene product of the W locus. Cell. 1990; 63(1):225-233. View reference

  7. 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. View reference

  8. Kondo M, Weissman IL, Akashi K. Identification of clonogenic common lymphoid progenitors in mouse bone marrow. Cell. 1997; 91(5):661-672. View reference

  9. Lian Z, Toki J, Yu C, et al. Intrathymically injected hemopoietic stem cells can differentiate into all lineage cells in the thymus: differences between c-kit+ cells and c-kit < low cells. Stem Cells. 1997; 15(6):430-436. View reference

  10. Mirmonsef P, Shelburne CP, Fitzhugh Yeatman C 2nd, Chong HJ, Ryan JJ. Inhibition of Kit expression by IL-4 and IL-10 in murine mast cells: role of STAT6 and phosphatidylinositol 3'-kinase. J Immunol. 1999; 163(5):2530-2539. View reference

  11. Ogawa M, Matsuzaki Y, Nishikawa S, et al. Expression and function of c-kit in hemopoietic progenitor cells. J Exp Med. 1991; 174(1):63-71. View reference

  12. Pulendran B, Lingappa J, Kennedy MK, et al. Developmental pathways of dendritic cells in vivo: distinct function, phenotype, and localization of dendritic cell subsets in FLT3 ligand-treated mice. J Immunol. 1997; 159(5):2222-2231. View reference

  13. Rodewald HR, Kretzschmar K, Swat W, Takeda S. Intrathymically expressed c-kit ligand (stem cell factor) is a major factor driving expansion of very immature thymocytes in vivo. Immunity. 1995; 3(3):313-319. View reference

  14. Valent P. The riddle of the mast cell: kit(CD117)-ligand as the missing link?. Immunol Today. 1994; 15(3):111-114. View reference

  15. Zsebo KM, Wypych J, McNiece IK, et al. Identification, purification, and biological characterization of hematopoietic stem cell factor from buffalo rat liver--conditioned medium. Cell. 1990; 63(1):195-201. View reference

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