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>   home   >   Products   >   Primary Antibodies   >   Immunology   >   Anti-Human CD161 Antibody, clone B199.2 (RPE)   

Anti-Human CD161 Antibody, clone B199.2

Mouse Anti-Human Monoclonal Antibody

     
  • FC - Anti-Human CD161 Antibody, clone B199.2  ABD12344
    Published customer image:Phycoerythrin conjugated Mouse anti Hyman CD161 antibody, clone B199.2 used for the evaluation of CD161 expression on transfected HEK-293 cells by flow cytometry.Image caption:Characterization of the ligand-binding domain of CD161/TT and CD161/CC receptors. (A) Fluorescence histograms obtained after incubation of 293-CD161/TT and -CD161/CC transfectants with control Ig-Fc (closed histograms) and LLT1-Fc or LLT1-Fc proteins multimerized by protein A (open histograms). Binding of the recombinant protein to CD161 was demonstrated by PE-conjugated goat anti-human IgG. (B) Dose-effect relationship of LLT1-Fc binding. 293-CD161/TT and -CD161/CC cells were incubated with increasing amounts of LLT1-Fc. Mean fluorescence intensity (MFI) was determined after a second incubation step with PE-conjugated goat anti-human IgG. Results represent mean values ± SEM calculated for three independent experiments; *p< 0.05 (Unpaired t-test). (C) Epitope-specific antibody competition in CD161/TT and -CC receptor variants. 293-CD161/TT and -CC cells were incubated with a fixed concentration (10 µg/ml) of PE-labelled anti-CD161 mAb B199.2 alone and in combination with increasing amounts of competing anti-CD161 mAb 191B8 or the anti-pig MHC class-II mAb MSA3 as IgG2a isotype control. Samples were analysed by flow cytometry and PE-MFI was determined. Competition is shown as % binding of mAb B199.2 and was calculated as follows: MFI (B199.2 plus 191B8) / MFI (B199.2 plus MSA3) x 100. MFI obtained with B199.2 in the absence of competing antibody was set as 100%. Results represent mean values ± SD calculated for five independent experiments; *pC Polymorphism in the Human KLRB1 Gene Alters Ligand Binding and Inhibitory Potential of CD161 Molecules.PLoS ONE 10: e0135682.
  • FC - Anti-Human CD161 Antibody, clone B199.2  ABD12344
    Published customer image:Mouse anti Hyman CD161 antibody, clone B199.2 used for the evaluation of CD161 expression on peripheral blood mononuclear cells by flow cytometry.Image caption:Assessment of lymphocyte subsets in individuals carrying different CD161 genotypes. PBMC were isolated and stained with the antibody combinations CD161/CD3/CD4 or CD161/CD3/CD56. Analyses were performed in gated viable lymphocytes as defined by forward- and side-scatter characteristics. (A) CD161 expression patterns on CD3- NK cells. Representative dot-plots obtained after staining of cells from one TT, TC, and CC individual are shown. Boxes indicate the CD56brightCD161+CD3- NK cell subset and the frequency. The numbers indicate % positive cells in each quadrant. (B) Decreased frequency of CD3-CD56+ NK cells in CC carriers. PBMC from 27 TT, 51 TC, and 16 carriers of CC were analyzed. Results are expressed as % NK cells among gated lymphocytes; the mean value calculated for each cohort is indicated by the horizontal line; *p< 0.05 . (C) Ratios of CD56dim to CD56bright NK cells. The frequencies of CD56dimCD161+CD3- and CD56brightCD161+CD3- NK cells were determined in gated lymphocytes from 22 TT, 46 TC, and 15 carriers of CC. The horizontal broken line is an arbitrary cut-off value. (D) Level of CD161 expression. Data are expressed as mean fluorescence intensity/MFI on gated CD3-CD56+ NK cells. (E) Proportion of NK cells co-expressing CD161. Data are expressed as % CD161+ cells in gated CD3-CD56+ NK cells. (F) CD161 expression patterns on CD3+ T cells. The numbers indicate % positive cells in each quadrant. (G) Proportion of CD4+ T cells co-expressing CD161. (H) Proportion of CD8+ T cells co-expressing CD161dim or CD161bright.From: Rother S, Hundrieser J, Pokoyski C, Kollrich S, Borns K, Blasczyk R, et al. The c.503T>C Polymorphism in the Human KLRB1 Gene Alters Ligand Binding and Inhibitory Potential of CD161 Molecules.PLoS ONE 10(8): e0135682.
  • WB - Anti-Human CD161 Antibody, clone B199.2  ABD12344
    Published customer image:Mouse anti Hyman CD161 antibody, clone B199.2 used for the evaluation of CD161 expression on transfected HEK-293 cells and peripheral blood mononuclear cells by western blotting and flow cytometry using phycoerythrin conjugated Mouse anti Human CD161 antibody clone B199.1 .Image caption:Characterization of 293 transfectants. 293 cells were “mock”-transfected with the empty pIRES2-AcGFP1 vector or vectors containing CD161 from a homozygous TT or CC individual. Analyses were performed after selection of stable transfectants. (A) Flow cytometry analysis of 293-mock, 293-CD161/TT, and 293-CD161/CC cells after staining with the PE-conjugated anti-CD161 mAb B199.2. Grey histograms represent staining with an isotype control mAb. The numbers represent mean fluorescence intensity obtained by anti-CD161 staining. (B) Detection of CD161 by Western-blotting. 293 transfectants were lysed, and protein lysates were separated by 9% SDS-PAGE. After transferring to PVDF membranes, samples were probed with the anti-CD161 mAb (B199.2). Binding of B199.2 was visualized by HRP-conjugated goat anti-mouse-IgG and ECL reagents. Protein loading was controlled by staining with a mAb against &alpha;-Actin. Representative results of at least two independent experiments are shown. Lysates of PBMC activated for five days with IL-12 were used as a further control to estimate the molecular weight of CD161 molecules.From: Rother S, Hundrieser J, Pokoyski C, Kollrich S, Borns K, Blasczyk R, et al. The c.503T>C Polymorphism in the Human KLRB1 Gene Alters Ligand Binding and Inhibitory Potential of CD161 Molecules.PLoS ONE 10(8): e0135682.
  • SPECIFICATION
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Product Information
Application
  • Applications Legend:
  • WB=Western Blot
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin-embedded Sections)
  • IHC-F=Immunohistochemistry (Frozen Sections)
  • IF=Immunofluorescence
  • FC=Flow Cytopmetry
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • E=ELISA
  • IP=Immunoprecipitation
  • DB=Dot Blot
  • CHIP=Chromatin Immunoprecipitation
  • FA=Fluorescence Assay
  • IEM=Immunoelectronmicroscopy
  • EIA=Enzyme Immunoassay
IHC-F, FC, IP
Primary Accession Q12918
Reactivity Human
Host Mouse
Clonality Monoclonal
Isotype IgG1
Clone Names B199.2
Calculated MW 25415 Da
Additional Information
Purification Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
Immunogen Purified human NK cells cultured in IL-2 (Bennettet al.1996)
Shelf Life 18 months from date of despatch.
Gene ID 3820
Other Names Killer cell lectin-like receptor subfamily B member 1, C-type lectin domain family 5 member B, HNKR-P1a, NKR-P1A, Natural killer cell surface protein P1A, CD161, KLRB1, CLEC5B, NKRP1A
Target/Specificity Mouse anti-Human CD161 antibody, clone B199.2 recognizes the human Killer cell lectin-like receptor subfamily B member 1, also known as CD161, C-type lectin domain family 5 member B, HNKR-P1a, NKR-P1A or Natural killer cell surface protein P1A. CD161 is a 225 amino acid ~25 kDa predicted molecular mass, single pass type II transmembrane glycoprotein with a single C-type lectin domain. CD161 is expressed by almost all NK cells and a subset of CD3+ve T cells (Lanier 1994).CD161, a member of the C-lectin is expressed as a disulphide bond-linked homodimeric cell surface protein, comprising two chains of ~40-44 kDa (Lanieret al.1994). CD161 acts as a receptor for another c-type lectin, LLT1 with roles in the regulation of NK cell and T cell function (Aldemiret al.2005).Mouse anti-Human CD161 antibody, clone B199.2 cross-competes with and recognizes a similar epitope to the DX1 monoclonal antibody (Lanieret al.1994).
Preservative & Stabilisers 0.09% Sodium Azide
Storage Store at +4℃ or at -20 ℃.
PrecautionsAnti-Human CD161 Antibody, clone B199.2 is for research use only and not for use in diagnostic or therapeutic procedures.
Research Areas
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References

1. Bennett, I.M. et al. (1996) Definition of natural killer NKR-PIA+/CD56-/CD16- functionally immature human NK cell subset that differentiates in vitro in the presence of interleukin 12.
J. Exp. Med. 184: 1845-1856. 2. Azzoni, L. et al. (1998) Differential transcriptional regulation of CD161 and a novel gene, 197/15a, by IL-2, IL-15, and IL-12 in NK and T cells.
J. Immunol. 161: 3493-3500. 3. de Lalla, C. et al. (2011) Invariant NKT Cell Reconstitution in Pediatric Leukemia Patients Given HLA-Haploidentical Stem Cell Transplantation Defines Distinct CD4+ and CD4- Subset Dynamics and Correlates with Remission State.
J Immunol. 186: 4490-9. 4. Huarte, E. et al. (2008) PILAR is a novel modulator of human T-cell expansion.
Blood. 112: 1259-68. 5. Williams, P.J. et al. (2009) Altered decidual leucocyte populations in the placental bed in pre-eclampsia and foetal growth restriction: a comparison with late normal pregnancy.
Reproduction. 138: 177-84. 6. Higai, K. et al. (2006) Binding of sialyl Lewis X antigen to lectin-like receptors on NK cells induces cytotoxicity and tyrosine phosphorylation of a 17-kDa protein.
Biochim Biophys Acta. 1760: 1355-63. 7. Birchall, M.A. et al. (2008) Immunologic response of the laryngeal mucosa to extraesophageal reflux.
Ann Otol Rhinol Laryngol. 117: 891-5. 8. Bossard, C. et al. (2012) Plasmacytoid dendritic cells and Th17 immune response contribution in gastrointestinal acute graft-versus-host disease.
Leukemia. 26: 1471-4. 9. Higai, K. et al. (2008) Binding of sialyl Lewis X antigen to lectin-like receptors on NK cells induces cytotoxicity and tyrosine phosphorylation of a 17-kDa protein
Biochim Biophys Acta. 1760: 1355-63. 10. Richter, J. et al. (2010) CD161 receptor participates in both impairing NK cell cytotoxicity and the response to glycans and vimentin in patients with rheumatoid arthritis.
Clin Immunol. 136: 139-47. 11. Abrahamsson, S.V. et al. (2013) Non-myeloablative autologous haematopoietic stem cell transplantation expands regulatory cells and depletes IL-17 producing mucosal-associated invariant T cells in multiple sclerosis.
Brain. 136: 2888-903. 12. Rother S et al. (2015) The c.503T>C Polymorphism in the Human KLRB1 Gene Alters Ligand Binding and Inhibitory Potential of CD161 Molecules.
PLoS One. 10 (8): e0135682.

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