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PE Rat Anti-Mouse CD117
PE Rat Anti-Mouse CD117

Two-color analysis of the expression of CD117 on mouse bone marrow cells. A single-cell suspension of BALB/c bone marrow was simultaneously stained with FITC-conjugated RA3-6B2 (anti-mouse CD45R/B220, Cat. No. 553087/553088, both panels) and PE-conjugated ACK45 (right panel) monoclonal antibodies. Flow cytometry was performed on a FACSCalibur™ (BDIS, San Jose, CA).

Two-color analysis of the expression of CD117 on mouse bone marrow cells. A single-cell suspension of BALB/c bone marrow was simultaneously stained with FITC-conjugated RA3-6B2 (anti-mouse CD45R/B220, Cat. No. 553087/553088, both panels) and PE-conjugated ACK45 (right panel) monoclonal antibodies. Flow cytometry was performed on a FACSCalibur™ (BDIS, San Jose, CA).

Product Details
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BD Pharmingen™
c-KIT; W; SCFR; Stem Cell Factor Receptor; Sl; Steel Factor Receptor; Ssm
Mouse (QC Testing)
Rat IgG2b, κ
Mouse bone marrow mast cells
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_395103
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

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

Recommended Assay Procedures

PE-conjugated anti-mouse CD117 mAb may be used in combination with the Mouse Lineage Panel (Cat. No. 559971) to identify hematopoietic stem cells.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  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.
553869 Rev. 7
Antibody Details
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ACK45

The ACK45 monoclonal antibody specifically binds to CD117 (c-Kit), a transmembrane tyrosine-kinase receptor which 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 Kitl gene (formerly steel, Sl), is a co-mitogen for hematopoietic stem cells and the myeloerythroid progenitors and a mast-cell differentiation factor. The Kit[W] and Kitl[Sl] 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 the CD90 (Thy-1)[lo] TER-119[-] CD45R/B220[-] CD11b (Mac-1)[-] Ly-6G (Gr-1)[-] CD4[-] CD8[-] Sca-1 (Ly-6A/E)[+] multipotent hematopoietic stem cells, progenitors committed to myeloid 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. The expression of CD117 has been included in protocols to isolate hematopoietic progenitors from fetal and adult peripheral blood. The specificity and functional activity of mAb ACK45 is very similar to that of the published antibody ACK22; it is reported to block the action of c-Kit in vitro and in vivo.

This antibody is routinely tested by flow cytometric analysis. Other applications were tested at BD Biosciences Pharmingen during antibody development only or reported in the literature.

553869 Rev. 7
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
553869 Rev.7
Citations & References
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Development References (20)

  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. (Biology). View Reference
  2. Austen KF, Boyce JA. Mast cell lineage development and phenotypic regulation. Leuk Res. 2001; 25(7):511-518. (Biology). 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. (Biology). View Reference
  4. Godfrey DI, Kennedy J, Mombaerts P, Tonegawa S, Zlotnik A. Onset of TCR-β gene rearrangement and role of TCR-β expression during CD3-CD4-CD8- thymocyte differentiation. J Immunol. 1994; 152(10):4783-4792. (Biology). View Reference
  5. Godfrey DI, Zlotnik A, Suda T. Phenotypic and functional characterization of c-kit expression during intrathymic T cell development. J Immunol. 1992; 149(7):2281-2285. (Biology). 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. (Biology). View Reference
  7. Katayama N, Shih JP, Nishikawa S, Kina T, Clark SC, Ogawa M. Stage-specific expression of c-kit protein by murine hematopoietic progenitors. Blood. 1993; 82(8):2353-2360. (Biology). View Reference
  8. Matsuzaki Y, Gyotoku J, Ogawa M, et al. Characterization of c-kit positive intrathymic stem cells that are restricted to lymphoid differentiation. J Exp Med. 1993; 178(4):1283-1292. (Biology). View Reference
  9. 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. (Biology). View Reference
  10. Moore T, Huang S, Terstappen LW, Bennett M, Kumar V. Expression of CD43 on murine and human pluripotent hematopoietic stem cells. J Immunol. 1994; 153(11):4978-4987. (Biology). View Reference
  11. Nishikawa S, Kusakabe M, Yoshinaga K, et al. In utero manipulation of coat color formation by a monoclonal anti-c-kit antibody: two distinct waves of c-kit-dependency during melanocyte development. EMBO J. 1991; 10(8):2111-2118. (Biology). View Reference
  12. 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. (Immunogen). View Reference
  13. Orlic D, Fischer R, Nishikawa S, Nienhuis AW, Bodine D. Purification and characterization of heterogeneous pluripotent hematopoietic stem cell populations expressing high levels of c-kit receptor. Blood. 1993; 82(3):762-770. (Biology). View Reference
  14. Rico-Vargas SA, Weiskopf B, Nishikawa S, Osmond DG. c-kit expression by B cell precursors in mouse bone marrow. Stimulation of B cell genesis by in vivo treatment with anti-c-kit antibody. J Immunol. 1994; 152(6):2845-2852. (Biology). View Reference
  15. Rodewald HR, Dessing M, Dvorak AM, Galli SJ. Identification of a committed precursor for the mast cell lineage. Science. 1996; 271(5250):818-822. (Biology). View Reference
  16. 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. (Biology). View Reference
  17. Rodewald HR, Kretzschmar K, Takeda S, Hohl C, Dessing M. Identification of pro-thymocytes in murine fetal blood: T lineage commitment can precede thymus colonization. EMBO J. 1994; 13(18):4229-4240. (Biology). View Reference
  18. Valent P. The riddle of the mast cell: kit(CD117)-ligand as the missing link?. Immunol Today. 1994; 15(3):111-114. (Biology). View Reference
  19. Yamamoto Y, Yasumizu R, Amou Y, et al. Characterization of peripheral blood stem cells in mice. Blood. 1996; 88(2):445-454. (Biology). View Reference
  20. Zsebo KM, Williams DA, Geissler EN, et al. Stem cell factor is encoded at the Sl locus of the mouse and is the ligand for the c-kit tyrosine kinase receptor. Cell. 1990; 63(1):213-224. (Biology). View Reference
View All (20) View Less
553869 Rev. 7

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