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Multiparameter flow cytometric analysis of CD161 expression on human peripheral blood leucocyte populations. Human peripheral blood was stained with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; Left Plot) or BD Horizon™ R718 Mouse Anti-Human CD161 antibody (Cat. No. 567078/567229; Right Plot). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Bivariate pseudocolor density plots showing the correlated expression of CD161 (or Ig Isotype control staining) versus side light scatter signals (SSC) were derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer and FlowJo™ software.
Multicolor flow cytometric analysis of CD161 expression on human peripheral blood lymphocytes. Human peripheral blood was stained with FITC Mouse Anti-Human CD56 antibody (Cat. No. 562794) and either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Left Plot) or BD Horizon™ R718 Mouse Anti-Human CD161 antibody (Right Plot). Erythrocytes were lysed with BD FACS™ Lysing Solution. Bivariate pseudocolor density plots showing the correlated expression of CD56 versus CD161 expression (or Ig Isotype control staining) were derived from events gated with forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer and FlowJo™ software.
BD Horizon™ R718 Mouse Anti-Human CD161
BD Horizon™ R718 Mouse Anti-Human CD161
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
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 CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
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Companion Products
The HP-3G10 monoclonal antibody specifically recognizes human CD161 which is also known as Natural killer cell surface protein P1A (NKR-P1A or NKRP1A) or C-type lectin domain family 5 member B (CLEC5B). CD161 is expressed on the cell surface as an 80 kDa disulfide-linked homodimeric type II transmembrane glycoprotein. It is encoded by KLRB1 (Killer cell lectin-like receptor subfamily B member 1) which belongs to the Ca2+-dependent C-type lectin superfamily. CD161 is expressed on NK cells and on subsets of CD4+ and CD8+ αβ T cells, NKT cells, γδ T cells, CD3+ thymocytes, and fetal liver cells. CD161 is preferentially expressed on memory/effector T cells. CD161 can reportedly inhibit NK cell-mediated cytotoxicity and IFN-γ production. Lectin-like transcript 1 (LLT-1), encoded by CLEC2D (C-type lectin domain family 2 member D), has been described as a ligand for CD161. LLT-1 is expressed on some activated dendritic cells (DC) and B cells.
The antibody was conjugated to BD Horizon Red 718, which has been developed exclusively by for BD Biosciences as a better alternative to Alexa Fluor™ 700. BD Horizon Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor™ 700.
Development References (4)
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Ida H, Morita C, Porcelli S, Anderson P. CD161 workshop: Reactivity of workshop natural killer cell monoclonal antibodies on fresh and interleukin 2-activated peripheral blood natural killer cells and CD4-negative CD8-negative αβ and γδ T-cell clones. 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.; 1997:313-317.
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Lanier LL, Chang C, Phillips JH. Human NKR-P1A. A disulfide-linked homodimer of the C-type lectin superfamily expressed by a subset of NK and T lymphocytes. J Immunol. 1994; 153(6):2417-2428. (Biology). View Reference
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Márquez C, Trigueros C, Franco JM, et al. Identification of a common developmental pathway for thymic natural killer cells and dendritic cells.. Blood. 1998; 91(8):2760-71. (Clone-specific). View Reference
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Poggi A, Revello V, Nanni L, Costa P, Moretta A. CD161 (human NKR-P1A) workshop panel 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.; 1997:307-312.
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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.