Two-parameter flow cytometric analysis of CD161 expression on human peripheral blood leucocyte populations. Human peripheral blood was stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714) or Alexa Fluor® 647 Mouse Anti-Human CD161 antibody (Cat. No. 566708). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-parameter flow-cytometric dot 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 viable leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Two-color 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 Alexa Fluor® 647 Mouse IgG1, κ Isotype Control or Alexa Fluor® 647 Mouse Anti-Human CD161 antibody (Cat. No. 566708). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-parameter flow-cytometric dot 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 viable human peripheral blood lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Two-parameter flow cytometric analysis of CD161 expression on human peripheral blood leucocyte populations. Human peripheral blood was stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714) or Alexa Fluor® 647 Mouse Anti-Human CD161 antibody (Cat. No. 566708). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-parameter flow-cytometric dot 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 viable leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Two-color 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 Alexa Fluor® 647 Mouse IgG1, κ Isotype Control or Alexa Fluor® 647 Mouse Anti-Human CD161 antibody (Cat. No. 566708). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-parameter flow-cytometric dot 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 viable human peripheral blood lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Product Details
BD Pharmingen™
CLEC5B; KLRB1; NKR-P1A; NKRP1A
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human NK cells
Flow cytometry (Routinely Tested)
5 µl
VI NK13
3820
AB_2869827
Aqueous buffered solution containing BSA and ≤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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
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.
- 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.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Lysing Solution 10X Concentrate CE_IVD
Size
100 mL
Cat No.
349202
Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO
Size
100 Tests
Cat No.
557714
FITC Mouse anti-Human CD56 (NCAM-1) RUO
Size
100 Tests
Cat No.
562794
566708 Rev. 1
Antibody Details
HP-3G10
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.
566708 Rev. 1
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-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.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
566708 Rev.1
Citations & References
Development References (5)
-
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: Flow cytometry). 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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Rosen DB, Cao W, Avery DT, et al. Functional consequences of interactions between human NKR-P1A and its ligand LLT1 expressed on activated dendritic cells and B cells.. J Immunol. 2008; 180(10):6508-17. (Biology). View Reference
566708 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.
