BD Horizon™ RB744 Mouse Anti-Human CD161 (KLRB1)
Clone HP-3G10 (RUO)
Multiparameter flow cytometric analysis of CD161 (KLRB1) expression on Human peripheral blood leukocyte populations. Human peripheral blood was stained with either BD Horizon™ RB744 Mouse IgG1, κ Isotype Control (Cat. No. 570519; Left Plot) or BD Horizon™ RB744 Mouse Anti-Human CD161 (KLRB1) antibody (Cat. No. 570612/570700; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD161 (KLRB1) [or Ig Isotype control staining] versus side light scatter signals (SSC-A) was derived from gated events with the forward and side light-scatter characteristics of intact leukocytes. 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 (KLRB1) expression on Human peripheral blood lymphocytes. Human peripheral blood was stained with PE Mouse Anti-Human CD56 (NCAM-1) antibody (Cat. No. 555516) and with either BD Horizon™ RB744 Mouse IgG1, κ Isotype Control (Cat. No. 570519; Left Plot) or BD Horizon™ RB744 Mouse Anti-Human CD161 (KLRB1) antibody (Cat. No. 570612/570700; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD161 (KLRB1) expression (or Ig Isotype control staining) versus CD56 (NCAM-1) was derived from gated events with forward and side light-scatter characteristics of intact peripheral blood lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
Multiparameter flow cytometric analysis of CD161 (KLRB1) expression on Human peripheral blood leukocyte populations. Human peripheral blood was stained with either BD Horizon™ RB744 Mouse IgG1, κ Isotype Control (Cat. No. 570519; Left Plot) or BD Horizon™ RB744 Mouse Anti-Human CD161 (KLRB1) antibody (Cat. No. 570612/570700; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD161 (KLRB1) [or Ig Isotype control staining] versus side light scatter signals (SSC-A) was derived from gated events with the forward and side light-scatter characteristics of intact leukocytes. 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 (KLRB1) expression on Human peripheral blood lymphocytes. Human peripheral blood was stained with PE Mouse Anti-Human CD56 (NCAM-1) antibody (Cat. No. 555516) and with either BD Horizon™ RB744 Mouse IgG1, κ Isotype Control (Cat. No. 570519; Left Plot) or BD Horizon™ RB744 Mouse Anti-Human CD161 (KLRB1) antibody (Cat. No. 570612/570700; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD161 (KLRB1) expression (or Ig Isotype control staining) versus CD56 (NCAM-1) was derived from gated events with forward and side light-scatter characteristics of intact 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 Horizon™
CLEC5B; KLRB1; NKR-P1A; NKRP1A
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human NK cells
Flow cytometry (Routinely Tested)
5 µl/test
VI NK13
3820
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 unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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.
Product Notices
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
RB744 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
570519
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
PE Mouse Anti-Human CD56 (NCAM-1) RUO
Size
100 Tests
Cat No.
555516
570700 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.
570700 Rev. 1
Format Details
RB744
The BD Horizon RealBlue™ 744 (RB744) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 746-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB744 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), we recommend using an optical filter centered near 750-nm (e.g., a 750/60-nm bandpass filter).
RB744
Blue 488 nm
498 nm
746 nm
570700 Rev.1
Citations & References
View product citations for antibody "570700" on CiteAb
Development References (5)
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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: 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
570700 Rev. 1
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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.