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      APC Hamster Anti-Mouse Notch4
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      569850Image1.png
      569850_569926-2.png

      Multicolor flow cytometric analysis of Notch4 expression on viable Mouse splenic dendritic cells.  C57BL/6 Mouse splenic leukocytes 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™ BV421 Hamster Anti-Mouse CD11c (Cat. No. 562782) and BD Horizon™ BUV395 Rat Anti-Mouse CD8a (Cat. No. 563786) antibodies and with either APC Hamster IgG1, κ Isotype Control (Cat. No. 553974; Left Plot) or APC Hamster Anti-Mouse Notch4 antibody (Cat. No. 569850/569926; Right Plot) at 1.0 μg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The contour plot showing the correlated expression of Notch4 (or Ig Isotype control staining) versus CD8a was derived from gated CD11c+ events with the forward and side light-scatter characteristics of viable (7-AAD-negative) leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

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      569850
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      BD Pharmingen™
      Notch 4; Notch gene homolog 4; Notch4
      Mouse (QC Testing)
      Armenian Hamster IgG, κ
      Mouse Extracellular Notch4 Domain Recombinant Protein
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      18132
      AB_3685369
      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 to the dye under optimum conditions and unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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      Recommended Assay Procedures

      BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation).  When fluorochrome conjugated antibodies are bound to BD® 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 BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      3. An isotype control should be used at the same concentration as the antibody of interest.
      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. This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
      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 http://regdocs.bd.com to access safety data sheets (SDS).
      8. For U.S. patents that may apply, see bd.com/patents.
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      Data Sheets

      DownloadTechnical Data Sheet (TDS)
      569850 Rev. 1
      Antibody Details
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      HMN4-14

      The HMN4-14 monoclonal antibody specifically recognizes an extracellular domain of mouse Neurogenic locus notch homolog protein 4 (Notch4 or Notch 4). Notch4 is a type 1 transmembrane glycoprotein receptor that is encoded by Notch4 (notch 4) which belongs to the Notch family containing four members, Notch1-Notch4. The Notch4 precursor is cleaved in the Golgi and presents as a cell surface heterodimeric receptor. The Notch4 receptor can bind to several membrane-bound ligands from the Delta-like or Jagged family. Upon ligand binding, Notch4 undergoes proteolytic cleavage that results in the release of the Notch intracellular domain, NICD. NICD translocates to the nucleus where it can form a transcriptional activator complex with various transcription factors. These multimeric complexes either positively or negatively regulate the expression of multiple genes including those that orchestrate many facets of embryonic development and the subsequent functioning of organ systems such as the hematopoietic, immune, nervous, and cardiovascular systems. Notch4 is reportedly expressed on CD8+ splenic dendritic cells, endothelial cells, and macrophages. It is weakly expressed on some CD4+CD8+ thymocytes but is not expressed on mature peripheral T cells.

      569850 Rev. 1
      Format Details
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      APC
      Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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      APC
      651 nm
      660 nm
      569850 Rev.1
      Citations & References
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      Development References (9)

      1. Gordon WR, Arnett KL, Blacklow SC. The molecular logic of Notch signaling--a structural and biochemical perspective.. J Cell Sci. 2008; 121(Pt 19):3109-19. (Biology). View Reference
      2. Koyanagi A, Sekine C, Yagita H. Expression of Notch receptors and ligands on immature and mature T cells.. Biochem Biophys Res Commun. 2012; 418(4):799-805. (Clone-specific: Flow cytometry). View Reference
      3. Moriyama Y, Sekine C, Koyanagi A, et al. Delta-like 1 is essential for the maintenance of marginal zone B cells in normal mice but not in autoimmune mice.. Int Immunol. 2008; 20(6):763-73. (Immunogen: Flow cytometry). View Reference
      4. Murata A, Yoshino M, Hikosaka M, et al. An evolutionary-conserved function of mammalian notch family members as cell adhesion molecules.. PLoS ONE. 2014; 9(9):e108535. (Clone-specific: Flow cytometry). View Reference
      5. Sauma D, Ramirez A, Alvarez K, Rosemblatt M, Bono MR. Notch signalling regulates cytokine production by CD8+ and CD4+ T cells.. Scand J Immunol. 2012; 75(4):389-400. (Clone-specific: Flow cytometry). View Reference
      6. Sekine C, Koyanagi A, Koyama N, Hozumi K, Chiba S, Yagita H. Differential regulation of osteoclastogenesis by Notch2/Delta-like 1 and Notch1/Jagged1 axes. Arthritis Res Ther. 2012; 14(2):R45-R58. (Clone-specific: Flow cytometry). View Reference
      7. Sekine C, Moriyama Y, Koyanagi A, et al. Differential regulation of splenic CD8- dendritic cells and marginal zone B cells by Notch ligands.. Int Immunol. 2009; 21(3):295-301. (Immunogen: Flow cytometry). View Reference
      8. Sekine C, Nanki T, Yagita H. Macrophage-derived delta-like protein 1 enhances interleukin-6 and matrix metalloproteinase 3 production by fibroblast-like synoviocytes in mice with collagen-induced arthritis.. Arthritis Rheumatol. 2014; 66(10):2751-61. (Clone-specific: Flow cytometry). View Reference
      9. Uyttendaele H, Marazzi G, Wu G, Yan Q, Sassoon D, Kitajewski J. Notch4/int-3, a mammary proto-oncogene, is an endothelial cell-specific mammalian Notch gene.. Development. 1996; 122(7):2251-9. (Biology). View Reference
      View All (9) View Less
      569850 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.

      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.