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      BUV496 Mouse Anti-Mouse NK-1.1
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      This product is the replacement for 741062.
      BUV496 Mouse Anti-Mouse NK-1.1
      Multicolor flow cytometric analysis of NK-1.1 expression on viable Mouse splenic leukocytes.  C57BL/6 Mouse splenocytes were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and 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™ BV711 Hamster Anti-Mouse CD3e antibody (Cat. No. 563123) and with either BD Horizon™ BUV496 Mouse IgG2a, κ Isotype Control (Cat. No. 612957; Left Plot) or BD Horizon™ BUV496 Mouse Anti-Mouse NK-1.1 antibody (Cat. No. 569716; 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 bivariate pseudocolor density plot showing the correlated expression of NK1.1 (or Ig Isotype control staining) versus CD3e was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) splenic 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.
      BUV496 Mouse Anti-Mouse NK-1.1
      Multicolor flow cytometric analysis of NK-1.1 expression on viable Mouse splenic leukocytes.  C57BL/6 Mouse splenocytes were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and 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™ BV711 Hamster Anti-Mouse CD3e antibody (Cat. No. 563123) and with either BD Horizon™ BUV496 Mouse IgG2a, κ Isotype Control (Cat. No. 612957; Left Plot) or BD Horizon™ BUV496 Mouse Anti-Mouse NK-1.1 antibody (Cat. No. 569716; 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 bivariate pseudocolor density plot showing the correlated expression of NK1.1 (or Ig Isotype control staining) versus CD3e was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) splenic 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.
      BUV496 Mouse Anti-Mouse NK-1.1 BUV496 Mouse Anti-Mouse NK-1.1
      Multicolor flow cytometric analysis of NK-1.1 expression on viable Mouse splenic leukocytes.  C57BL/6 Mouse splenocytes were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and 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™ BV711 Hamster Anti-Mouse CD3e antibody (Cat. No. 563123) and with either BD Horizon™ BUV496 Mouse IgG2a, κ Isotype Control (Cat. No. 612957; Left Plot) or BD Horizon™ BUV496 Mouse Anti-Mouse NK-1.1 antibody (Cat. No. 569716; 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 bivariate pseudocolor density plot showing the correlated expression of NK1.1 (or Ig Isotype control staining) versus CD3e was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) splenic 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.
      Multicolor flow cytometric analysis of NK-1.1 expression on viable Mouse splenic leukocytes.  C57BL/6 Mouse splenocytes were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and 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™ BV711 Hamster Anti-Mouse CD3e antibody (Cat. No. 563123) and with either BD Horizon™ BUV496 Mouse IgG2a, κ Isotype Control (Cat. No. 612957; Left Plot) or BD Horizon™ BUV496 Mouse Anti-Mouse NK-1.1 antibody (Cat. No. 569716; 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 bivariate pseudocolor density plot showing the correlated expression of NK1.1 (or Ig Isotype control staining) versus CD3e was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) splenic 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.
      Product Details
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      BD Horizon™
      Klrb1b, CD161b, Nkrp1b; Klrb1c, CD161c, NK1.1, Nkrp1c
      Mouse (QC Testing)
      Mouse C3H x BALB/c IgG2a, κ
      Mouse NK-1+ Spleen and Bone Marrow Cells
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      17059, 80782
      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 cells and BD® CompBeads 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 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).

      Note:  When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed.  For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).

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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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      7. For U.S. patents that may apply, see bd.com/patents.
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      569716 Rev. 1
      Antibody Details
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      PK136

      In the mouse, at least three members of the Klrb (Killer cell lectin-like receptor, subfamily b; formerly NKR-P1) gene family have been identified (Klrb1a/NKR-P1A, Klrb1b/NKR-P1B, and Klrb1c/NKR-P1C); but in the human gene family, a single homologue has been designated KLRB1, NKR-P1A, or CD161. The KLRB1/NKR-P1 family of proteins are type-II-transmembrane C-type lectin receptors. KLRB1C/NKR-P1C activates NK-cell cytotoxicity, while KLRB1B/NKR-P1B functions as an inhibitory receptor. KLRB1B/NKR-P1B protein has intracellular Immunoreceptor Tyrosine-based Inhibitory Motif (ITIM), while KLRB1C/NKR-P1C lacks ITIM and activates via association with Fc Receptor γ chain. Strikingly, KLRB1B/NKR-P1B and KLRB1C/NKR-P1C share 96% amino acid sequence identity in their extracellular C-type lectin domains. The PK136 antibody reacts with the NK-1.1 surface antigen (CD161c) encoded by the Klrb1c/NKR-P1C gene expressed on natural killer (NK) cells in selected strains of mice (eg, C57BL, FVB/N, NZB, but not A, AKR, BALB/c, CBA/J, C3H, C57BR, C58, DBA/1, DBA/2, NOD, SJL, 129) and the CD161b antigen encoded by the Klrb1b/NKR-P1B gene expressed only on Swiss NIH and SJL mice, but not on C57BL/6. Expression of KLRB1C/NKR-P1C protein is correlated with the ability to lyse tumor cells in vitro and to mediate rejection of bone marrow allografts. The NK-1.1 marker is useful in defining NK cells; however, the antigen is also expressed on a rare, specialized population of T lymphocytes (NK-T cells) and some cultured monocytes. Plate-bound PK136 mAb, in combination with low concentrations of IL-2, induces proliferation of a subset of NK cells.

      569716 Rev. 1
      Format Details
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      BUV496
      The BD Horizon Brilliant™ Ultraviolet 496 (BUV496) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This tandem fluorochrome is comprised of a BUV395 donor with an excitation maximum (Ex Max) of 350-nm and an acceptor dye with an emission maximum (Em Max) at 496-nm. BUV496, driven by BD innovation, is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 500-nm (e.g., 515/30-nm bandpass filter). The acceptor dye can be excited by the Violet (405-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.
      altImg
      BUV496
      Ultraviolet 355 nm
      350 nm
      496 nm
      569716 Rev.1
      Citations & References
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      Development References (11)

      1. Arase N, Arase H, Park SY, Ohno H, Ra C, Saito T. Association with FcRgamma is essential for activation signal through NKR-P1 (CD161) in natural killer (NK) cells and NK1.1+ T cells. J Exp Med. 1997; 186(12):1957-1963. (Clone-specific: Cytotoxicity, Flow cytometry, Fluorescence activated cell sorting, Functional assay, Immunofluorescence, Immunoprecipitation, Stimulation). View Reference
      2. Carlyle JR, Martin A, Mehra A, Attisano L, Tsui FW, Zuniga-Pflucker JC. Mouse NKR-P1B, a novel NK1.1 antigen with inhibitory function. J Immunol. 1999; 162(10):5917-5923. (Clone-specific: Cytotoxicity, Flow cytometry, Immunoprecipitation, Inhibition, Stimulation). View Reference
      3. Giorda R, Trucco M. Mouse NKR-P1. A family of genes selectively coexpressed in adherent lymphokine-activated killer cells. J Immunol. 1991; 147(5):1701-1708. (Biology). View Reference
      4. Koo GC, Peppard JR. Establishment of monoclonal anti-Nk-1.1 antibody. Hybridoma. 1984; 3(3):301-303. (Immunogen: Cytotoxicity, Flow cytometry). View Reference
      5. Kung SK, Su RC, Shannon J, Miller RG. The NKR-P1B gene product is an inhibitory receptor on SJL/J NK cells. J Immunol. 1999; 162(10):5876-5887. (Clone-specific: Activation, Calcium Flux, Flow cytometry, Fluorescence activated cell sorting, Immunoprecipitation, Inhibition, Stimulation). View Reference
      6. Lanier LL. Natural killer cells: from no receptors to too many. Immunity. 1997; 6(4):371-378. (Biology). View Reference
      7. Reichlin A, Yokoyama WM. Natural killer cell proliferation induced by anti-NK1.1 and IL-2. Immunol Cell Biol. 1998; 76(2):143-152. (Clone-specific: Activation, Cytotoxicity, Functional assay, Stimulation). View Reference
      8. Sentman CL, Kumar V, Koo G, Bennett M. Effector cell expression of NK1.1, a murine natural killer cell-specific molecule, and ability of mice to reject bone marrow allografts. J Immunol. 1989; 142(6):1847-1853. (Clone-specific: Flow cytometry, Immunofluorescence). View Reference
      9. Vicari AP, Zlotnik A. Mouse NK1.1+ T cells: a new family of T cells. Immunol Today. 1996; 17(2):71-76. (Biology). View Reference
      10. Yokoyama WM, Seaman WE. The Ly-49 and NKR-P1 gene families encoding lectin-like receptors on natural killer cells: the NK gene complex. Annu Rev Immunol. 1993; 11:613-635. (Biology). View Reference
      11. Yu YY, Kumar V, Bennett M. Murine natural killer cells and marrow graft rejection. Annu Rev Immunol. 1992; 10:189-213. (Biology). View Reference
      View All (11) View Less
      569716 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.

      For Research Use Only. Not for use in diagnostic or therapeutic procedures.

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