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    BB700 Mouse Anti-Human CD158b
    Product Details
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    BD OptiBuild™
    CD158b1/KIR2DL2/NKAT-6; CD158b2/KIR2DL3/NKAT-2; CD158j/KIR2DS2/NKAT-5
    Human (Tested in Development)
    Mouse BALB/c IgG2b, κ
    Human NK Cells
    Flow cytometry (Qualified)
    0.2 mg/ml
    VI NK8
    AB_2743574
    Aqueous buffered solution containing ≤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 BB700 under optimal conditions that minimize unconjugated dye and antibody.
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    Recommended Assay Procedures

    For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794 or 566349).

    When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values.

    For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

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    Product Notices

    1. This antibody was developed for use in flow cytometry.
    2. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
    3. Researchers should determine the optimal concentration of this reagent for their individual applications.
    4. An isotype control should be used at the same concentration as the antibody of interest.
    5. 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.
    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 www.bdbiosciences.com/us/s/resources for technical protocols.
    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. BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
    10. Cy is a trademark of GE Healthcare.
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    746236 Rev. 1
    Antibody Details
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    CH-L

    The CH-L monoclonal antibody specifically binds to CD158b proteins. These proteins are 50-58 kDa type I glycoproteins that belong to the Killer cell immunoglobulin-like receptor (KIR) family: (KIR2DL2/L3/S2). They are also known as CD158b1 (KIR2DL2; NKAT-6; p58.2), CD158b2 (KIR2DL3; NKAT-2; p58.2), or CD158j (KIR2DS2; NKAT-5; p50.2). The CD158b molecules are composed of two extracellular Ig-like domains, and a transmembrane region. CD158b1 and CD158b2 also possess long (84 or 76 amino acids, respectively) cytoplasmic tails with two immunoreceptor tyrosine-based inhibition motifs (ITIM) whereas CD158j has a short (39 amino acid) cytoplasmic tail that lacks the ITIM motif.  CD158b molecules are expressed on NK cells and subsets of TCR αβ+ cells or TCR γδ+ cells. Ligand- or CH-L antibody-bound CD158b1 or CD158b2 can reportedly inhibit cytolytic NK and T cell responses to various stimuli including certain target cells expressing MHC class I ligands encoded by HLA-C alleles (Cw 1, 3, 7 and 8).  CD158j reportedly can enhance some cellular cytolytic responses.

    The antibody was conjugated to BD Horizon™ BB700, which is part of the BD Horizon Brilliant™ Blue family of dyes.   It is a polymer-based tandem dye developed exclusively by BD Biosciences.  With an excitation max of 485 nm and an emission max of 693 nm, BD Horizon BB700 can be excited by the 488 nm laser and detected in a standard PerCP-Cy™5.5 set (eg, 695/40-nm filter). This dye provides a much brighter alternative to PerCP-Cy5.5 with less cross laser excitation off the 405 nm and 355 nm lasers.

    746236 Rev. 1
    Format Details
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    BB700
    The BD Horizon Brilliant™ Blue 700 (BB700) Dye is part of the BD Horizon Brilliant™ Blue family of dyes. This tandem fluorochrome is comprised of a polymer-technology dye donor with an excitation maximum (Ex Max) of 476-nm and an acceptor dye with an emission maximum (Em Max) at 695-nm. Driven by BD innovation, BB700 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 695-nm (e.g., a 695/20-nm bandpass filter). The donor dye can be excited by the Violet (405 nm) laser and the acceptor dye can be excited by the red (627–640 nm) laser resulting in cross-laser excitation and fluorescence spillover. BB700 Reagents are significantly brighter than equivalent PerCP or PerCP-Cy5.5 reagents and are less sensitive to photobleaching. In addition, BB700 shows much less excitation by the violet (407-nm) laser resulting in less spillover. BB700 has minimal yellow green (562-nm) excitation and is ideal for instruments with both blue (488-nm) and yellow green (562-nm) lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
    altImg
    BB700
    Blue 488 nm
    476 nm
    695 nm
    746236 Rev.1
    Citations & References
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    Development References (8)

    1. Cambiaggi A, Orengo AM, Meazza R, et al. The natural killer-related receptor for HLA-C expressed on T cells from CD3+ lymphoproliferative disease of granular lymphocytes displays either inhibitory or stimulatory function. Blood. 1996; 87(6):2369-2375. (Biology). View Reference
    2. Colonna M, Samaridis J. Cloning of immunoglobulin-superfamily members associated with HLA-C and HLA-B recognition by human natural killer cells. Science. 1995; 268(5209):405-408. (Biology). View Reference
    3. Ferrini S, Cambiaggi A, Meazza R, et al. T cell clones expressing the natural killer cell-related p58 receptor molecule display heterogeneity in phenotypic properties and p58 function. Eur J Immunol. 1994; 24(10):2294-2298. (Clone-specific: Blocking, Cell separation, Flow cytometry, Functional assay, Inhibition, Radioimmunoassay, Western blot). View Reference
    4. Kim J, Chwae YJ, Kim MY, Choi IH, Park JH, Kim SJ. Molecular basis of HLA-C recognition by p58 natural killer cell inhibitory receptors. J Immunol. 1997; 159(8):3875-3882. (Biology). View Reference
    5. Moretta A, Bottino C, Biassoni R. CD158a (p58.1/p50.1) and CD158b (p58.2/p50.2) natural killer receptors for HLA-C alleles: Workshop Report. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:290-292.
    6. Pascal V, Vivier E, Andre P. CD158 (killer immunoglobulin-like receptors family) report. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:412-413.
    7. Warren HS, Kinnear BF. CD158a and b Workshop: Killer-inhibitory receptors and natural killer cell proliferation. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:292-294.
    8. van Bergen J, Thompson A, van der Slik A, Ottenhoff TH, Gussekloo J, Koning F. Phenotypic and functional characterization of CD4 T cells expressing killer Ig-like receptors. J Immunol. 2004; 173(11):6719-6726. (Clone-specific: Flow cytometry). View Reference
    View All (8) View Less
    746236 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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