CD161 antibody | B199.2

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Mouse anti Human CD161:StarBright UltraViolet 700

Product Type: Monoclonal Antibody
Clone: B199.2
Isotype: IgG1
Specificity: CD161

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MCA1855SBUV700 Datasheet SDS SDS	F *	100 Tests/0.5ml		Log in
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Anti Human CD161 Antibody, clone B199.2 gallery image 1 — Figure A. RPE-Alexa Fluor^® 750 conjugated Mouse anti Human CD3 antibody, clone UCHT1 (**MCA463P750**) and Alexa Fluor^® 647 conjugated Mouse IgG1 isotype control (**MCA928A647**).
Figure B. RPE-Alexa Fluor^® 750 conjugated Mouse anti Human CD3 antibody, clone UCHT1 (**MCA463P750**) and Alexa Fluor^® 647 conjugated Mouse anti Human CD161 antibbody, clone B199.2 (**MCA1855A647**). All experiments performed on red cell lysed human blood gated on lymphocytes in the presence of Human Seroblock (**BUF070A**).
Data acquired on the **ZE5 Cell Analyzer**.

Anti Human CD161 Antibody, clone B199.2 gallery image 2 — **Published customer image:**
Mouse anti Human CD161 antibody, clone B199.2 (**MCA1855**) used for the evaluation of CD161 expression on human 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 (ANOVA). (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*. (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**.
This image is from an open access article distributed under the terms of the Creative Commons Attribution License.

Anti Human CD161 Antibody, clone B199.2 gallery image 3 — **Published customer image:**
Mouse anti Hyman CD161 antibody, clone B199.2 (**MCA1855**) 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 (**MCA1855PE**).
**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 α-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*. (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**.
This image is from an open access article distributed under the terms of the Creative Commons Attribution License.

Anti Human CD161 Antibody, clone B199.2 gallery image 4 — **Published customer image:**
Phycoerythrin conjugated Mouse anti Human CD161 antibody, clone B199.2 (**MCA1855PE**) 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 (IgG2a) 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; *p<0.05 (Unpaired t-test).

From: Rother S, Hundrieser J, Pokoyski C, Kollrich S, Borns K, Blasczyk R, *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**.
This image is from an open access article distributed under the terms of the Creative Commons Attribution License.

Anti Human CD161 Antibody, clone B199.2 gallery image 5 — **Published customer image:**
Mouse anti Human CD161 antibody, clone B199.2 (**MCA1855G**) used for the demonstration of CD161 expression in human tonsil by immunohistochemistry on formalin fixed, paraffin embedded tissue sections.
**Image caption:**
CD161 is expressed in the germinal center environment.
Staining with anti-CD161 antibody (brown; B199.2 clone) of FFPE human tonsillar tissue (CD161 20x, Isotype control 10x) revealed an expression pattern similar to the one of FDCs. Representative images of one out of 3 independent experiments are shown.

From: Llibre A, López-Macías C, Marafioti T, Mehta H, Partridge A, Kanzig C, Rivellese F, Galson JD, Walker LJ, Milne P, Phillips RE, Kelly DF, Freeman GJ, El Shikh ME, Klenerman P, Willberg CB.
LLT1 and CD161 Expression in Human Germinal Centers Promotes B Cell Activation and CXCR4 Downregulation.
**J Immunol. 2016 Mar 1;196(5):2085-94**.
This image is from an open access article distributed under the terms of the Creative Commons Attribution License.

Anti Human CD161 Antibody, clone B199.2 gallery image 6 — **Published customer image:**
Mouse anti Human CD161 antibody, clone B199.2 (**MCA1855G**) used for the demonstration of CD161 expression in human tonsil by immunofluorescence
**Image caption:**
Immunofluorescent staining of frozen human tonsils with 35 the FDC marker and anti-CD161 antibody clone B199.2 revealed expression of CD161 on FDCs.

From: Llibre A, López-Macías C, Marafioti T, Mehta H, Partridge A, Kanzig C, Rivellese F, Galson JD, Walker LJ, Milne P, Phillips RE, Kelly DF, Freeman GJ, El Shikh ME, Klenerman P, Willberg CB.
LLT1 and CD161 Expression in Human Germinal Centers Promotes B Cell Activation and CXCR4 Downregulation.
**J Immunol. 2016 Mar 1;196(5):2085-94**.
This image is from an open access article distributed under the terms of the Creative Commons Attribution License.

Anti Human CD161 Antibody, clone B199.2 gallery image 7 — Human whole blood was stained with Anti-Human CD3 StarBright Blue 810 (**MCA463SBB810**) only (Figure A) or with with Anti-Human CD3 StarBright Blue 810 (**MCA463SBB810**) and Anti-Human CD161 StarBright UltraViolet 540 (**MCA1855SBUV540**) (Figure B). Experiments were performed on isolated red-cell lysed blood, gated on live, single Lymphocytes. Data were acquired on a Cytek Aurora.

Anti Human CD161 Antibody, clone B199.2 gallery image 8 — Human whole blood was stained with Anti-Human CD3 StarBright Blue 810 (**MCA463SBB810**) only (Figure A) or with with Anti-Human CD3 StarBright Blue 810 (**MCA463SBB810**) and Anti-Human CD161 StarBright UltraViolet 700 (**MCA1855SBUV700**) (Figure B). Experiments were performed on isolated red-cell lysed blood, gated on live, single Lymphocytes. Data were acquired on a Cytek Aurora.

Anti Human CD161 Antibody, clone B199.2 gallery image 9 — Human whole blood was stained with Anti-Human CD3 StarBright UltraViolet 400 (**MCA463SBUV400**) only (Figure A) or with with Anti-Human CD3 StarBright UltraViolet 400 (**MCA463SBUV400**) and Anti-Human CD161 StarBright UltraViolet 770 (MCA1855SBUV770) (Figure B). Experiments were performed on isolated red-cell lysed blood, gated on live, single Lymphocytes. Data were acquired on a Cytek Aurora.

Anti Human CD161 Antibody, clone B199.2 gallery image 10 — Human whole blood was stained with Anti-Human CD3 StarBright Violet 790 (**MCA463SBV790**) only (Figure A) or with with Anti-Human CD3 StarBright Violet 790 (**MCA463SBV790**) and Anti-Human CD161 StarBright Violet 535 (**MCA1855SBV535**) (Figure B). Experiments were performed on isolated red-cell lysed blood, gated on live, single Lymphocytes. Data were acquired on a Cytek Aurora.

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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 (Lanier et 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 (Aldemir et al. 2005).

Mouse anti Human CD161 antibody, clone B199.2 cross-competes with and recognizes a similar epitope to the DX1 monoclonal antibody (Lanier et al. 1994).

Target Species: Human
Product Form: Purified IgG conjugated to StarBright UltraViolet 700 - liquid
Preparation: Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
Buffer Solution: Phosphate buffered saline
Preservative Stabilisers: 0.09% Sodium Azide (NaN₃)
1% Bovine Serum Albumin
0.1% Pluronic F68
0.1% PEG 3350
0.05% Tween 20
Immunogen: Purified human NK cells cultured in IL-2 (Bennett et al. 1996)
Approx. Protein Concentrations: For information on the concentration of our StarBright Dye conjugated reagents please visit our FAQ page.
Fusion Partners: Spleen cells from immunised BALB/c mice were fused with cells of the mouse P2X63.Ag8.653 myeloma cell line.
Max Ex/Em: Fluorophore Excitation Max (nm) Emission Max (nm)

StarBright UltraViolet 700 340 700
Regulatory: For research purposes only
Guarantee: 12 months from date of despatch
Acknowledgements: This product is covered by U.S. Patent No. 10,150,841 and related U.S. and foreign counterparts

Store at +4°C. DO NOT FREEZE.
This product should be stored undiluted.

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

Application Name	Verified	Min Dilution	Max Dilution
Flow Cytometry ¹		Neat

¹Staining at lower temperatures generally requires longer incubation to reach antibody binding equilibrium, and this effect is more pronounced for StarBright Dyes due to their larger size. For optimum staining, Bio-Rad recommends incubating samples either overnight at 4°C or 1 hour at room temperature.

Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.

Flow Cytometry: Use 5μl of the suggested working dilution to label 0.5x10⁶ cells in 100μl. Best practices suggest a 5 min centrifugation at 6,000g prior to sample application.

Description	Product Code	Applications	Pack Size	Your Price
Human Seroblock	BUF070A	F	50 Test	Log in
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Description	Human Seroblock
Human Seroblock	BUF070B	F	200 Test	Log in
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Description	Human Seroblock

References for CD161 antibody

Bennett, I.M. et al. (1996) Definition of a natural killer NKR-P1A+/CD56-/CD16- functionally immature human NK cell subset that differentiates in vitro in the presence of interleukin 12.
J Exp Med. 184 (5): 1845-56.
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 (7): 3493-500.
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 (9): 1355-63.
Birchall, M.A. et al. (2008) Immunologic response of the laryngeal mucosa to extraesophageal reflux.
Ann Otol Rhinol Laryngol. 117: 891-5.
Huarte, E. et al. (2008) PILAR is a novel modulator of human T-cell expansion.
Blood. 112: 1259-68.
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.
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.
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.

View The Latest Product References

Bossard, C. et al. (2012) Plasmacytoid dendritic cells and Th17 immune response contribution in gastrointestinal acute graft-versus-host disease.
Leukemia. 26: 1471-4.
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.
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.

Flow Cytometry
Immunohistology - Frozen

Synonyms: NKR-P1
UniProt: Q12918
Entrez Gene: KLRB1
GO Terms: GO:0005886 plasma membrane; GO:0016021 integral to membrane; GO:0004888 transmembrane receptor activity; GO:0005529 sugar binding; GO:0007166 cell surface receptor linked signaling pathway