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Two-color flow cytometric analysis of CD117 (c-kit) expression on mouse bone marrow cells. C57BL/6 mouse bone marrow cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with BD Horizon™ BUV395 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 563793) and with either BD Horizon™ BV421 Rat IgG2b, κ Isotype Control (Cat. No. 562603; Left Plot) or BD Horizon™ BV421 Rat Anti-Mouse CD117 (c-kit) antibody (Cat. No. 567818; Right Plot) at 0.5 μg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The pseudocolor density plot showing the correlated expression of CD117 (c-Kit) [or Ig Isotype control staining)] versus CD45R/B220 was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown in this Technical Data Sheet are not lot specific.
BD Horizon™ BV421 Rat Anti-Mouse CD117 (c-kit)
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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 cells and CompBead to ensure that BD® CompBeads are appropriate for your specific cellular application.
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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- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- 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.
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The ACK2 monoclonal antibody specifically binds to CD117, which is also known as Proto-oncogene c-Kit (c-kit), Stem Cell Factor Receptor (SCFR), and Mast/stem cell growth factor receptor. This ~145 kDa type I transmembrane glycoprotein is encoded by Kit (KIT proto-oncogene receptor tyrosine kinase) which belongs to the receptor tyrosine kinase family (RTK) within the immunoglobulin superfamily (IgSF). CD117 (c-kit) serves as a receptor for c-Kit ligand (aka, stem cell factor or SCF). This receptor is expressed on multipotent hematopoietic stem cells and progenitors committed to myeloid and/or erythroid lineages, and precursors of B and T lymphocytes. CD117 (c-kit)-mediated signaling plays an important role in the activation, growth, proliferation, differentiation, and survival of cells in sustaining hematopoiesis. It is also involved in the development of mast cells and melanocytes. The specificity and functional activity of the ACK2 antibody is like that of another CD117 (c-kit)-specific antibody, ACK45. ACK2 is reported to block CD117 (c-kit) signaling in vitro and in vivo.
研发参考 (6)
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Czechowicz A, Kraft D, Weissman IL, Bhattacharya D. Efficient transplantation via antibody-based clearance of hematopoietic stem cell niches.. Science. 2007; 318(5854):1296-9. (Clone-specific: Flow cytometry, Functional assay, Inhibition, In vivo exacerbation). 查看参考
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Géraud C, Koch PS, Zierow J, et al. GATA4-dependent organ-specific endothelial differentiation controls liver development and embryonic hematopoiesis.. J Clin Invest. 2017; 127(3):1099-1114. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). 查看参考
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Hozumi K, Kobori A, Sato T, et al. Pro-T cells in fetal thymus express c-kit and RAG-2 but do not rearrange the gene encoding the T cell receptor beta chain.. Eur J Immunol. 1994; 24(6):1339-44. (Clone-specific: Functional assay, Inhibition). 查看参考
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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. (Immunogen: Functional assay, Immunohistochemistry, Inhibition, In vivo exacerbation). 查看参考
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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: Functional assay, Inhibition, In vivo exacerbation). 查看参考
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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. (Clone-specific: Immunofluorescence, In vivo exacerbation). 查看参考
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