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mCD117_BV711_CD45R_APC.png

Multicolor flow cytometric analysis of CD117 expression on mouse bone marrow cells. Bone marrow cells from BALB/c mice were preincubated with Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) (Cat. no. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD45R/B220 (Cat. No. 553092/561880) and with either BD Horizon™ BV711 Rat IgG2b, κ Isotype Control (Cat. No. 563045, Left Panel) or with BD Horizon™ BV711 Rat Anti-Mouse CD117 antibody (Cat. No. 563160, Right Panel). Two-color flow cytometric dot plots showing the expression of CD117 (or Ig isotype control staining) versus CD45R/B220 were derived from gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

563160Image1.png

Multicolor flow cytometric analysis of CD117 expression on mouse bone marrow cells. Bone marrow cells from BALB/c mice were preincubated with Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) (Cat. no. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD45R/B220 (Cat. No. 553092/561880) and with either BD Horizon™ BV711 Rat IgG2b, κ Isotype Control (Cat. No. 563045, Left Panel) or with BD Horizon™ BV711 Rat Anti-Mouse CD117 antibody (Cat. No. 563160, Right Panel). Two-color flow cytometric dot plots showing the expression of CD117 (or Ig isotype control staining) versus CD45R/B220 were derived from gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

mCD117_BV711_CD45R_APC.png 563160Image1.png Red-Cap_Violet.png

Multicolor flow cytometric analysis of CD117 expression on mouse bone marrow cells. Bone marrow cells from BALB/c mice were preincubated with Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) (Cat. no. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD45R/B220 (Cat. No. 553092/561880) and with either BD Horizon™ BV711 Rat IgG2b, κ Isotype Control (Cat. No. 563045, Left Panel) or with BD Horizon™ BV711 Rat Anti-Mouse CD117 antibody (Cat. No. 563160, Right Panel). Two-color flow cytometric dot plots showing the expression of CD117 (or Ig isotype control staining) versus CD45R/B220 were derived from gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Multicolor flow cytometric analysis of CD117 expression on mouse bone marrow cells. Bone marrow cells from BALB/c mice were preincubated with Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) (Cat. no. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD45R/B220 (Cat. No. 553092/561880) and with either BD Horizon™ BV711 Rat IgG2b, κ Isotype Control (Cat. No. 563045, Left Panel) or with BD Horizon™ BV711 Rat Anti-Mouse CD117 antibody (Cat. No. 563160, Right Panel). Two-color flow cytometric dot plots showing the expression of CD117 (or Ig isotype control staining) versus CD45R/B220 were derived from gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Product Details
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BD Horizon™
c-KIT; W; SCFR; Stem Cell Factor Receptor; Sl; Steel Factor Receptor; Ssm
Mouse (QC Testing)
Rat WI, also known as Wistar (outbred) IgG2b, κ
Mouse Bone Marrow Mast Cells
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2722510
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 with BD Horizon™ BV711 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV711 were removed.
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Recommended Assay Procedures

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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Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  7. BD Horizon Brilliant Violet 711 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
  8. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  9. Cy is a trademark of GE Healthcare.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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563160 Rev. 2
Antibody Details
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2B8

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.

563160 Rev. 2
Format Details
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BV711
The BD Horizon Brilliant Violet™ 711 (BV711) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 407-nm and an acceptor dye with an emission maximum (Em Max) at 713-nm. BV711, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 710-nm (e.g., a 712/20-nm bandpass filter). The acceptor dye can be excited by the Red (628–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV711
407 nm
713 nm
563160 Rev.2
Citations & References
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View product citations for antibody "563160" on CiteAb

Development References (13)

  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. 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. (Clone-specific: Flow cytometry). 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. 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. (Immunogen: Flow cytometry, Immunoprecipitation). 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. (Biology). 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. (Biology). View Reference
  10. 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. (Biology). View Reference
  11. 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
  12. 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
  13. 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. (Biology). View Reference
View All (13) View Less
563160 Rev. 2

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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.