CD161 antibody

Catalog No. ABIN487476

Product Details anti-CD161 Antibody

Target: CD161 (KLRB1) (Killer Cell Lectin-Like Receptor Subfamily B, Member 1 (KLRB1))

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This CD161 antibody is un-conjugated

Application: Flow Cytometry (FACS)

Specificity: This antibody reacts with Human CD161 antigen.

Cross-Reactivity (Details): Species reactivity (tested):Human.

Characteristics: Synonyms: HNKR-P1a, CLEC5B, NKRP1A, Killer cell lectin-like receptor subfamily B member 1, Naturalkiller cell surface protein P1A, C-type lectin domain family 5 member B

Purification: Protein-A Sepharose Chromatography.

Immunogen: Human polyclonal NK cells. Remarks: Hybridoma was established by fusion of mouse myeloma cell SP2 with Balb/cmouse splenocyte.

Clone: HP-3G10

Isotype: IgG1

Application Details

Application Notes: Flow Cytometry: 10-20 μg/mL (final concentration). Positive Control: Lymphocyte cells. Detailed procedure is provided in Protocols.
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.

Protocol: Flow Cytometric analysis for floating cellsProtocol 1We usually use Fisher tubes or equivalents as reaction tubes for all step described below. 1) Resuspend the cells with washing buffer (5x10e6 cells/mL). 2) Add 50 µL of the cell suspension into each tube, and centrifuge at 500xg for 1 minute atroom temperature (20~25°C). Remove supernatant by careful aspiration. 3) Add 10 µL of normal goat serum containing 1 mg/mL normal human IgG and 0. 1% NaN3to the cell pellet after tapping. Mix well, and incubate for 5 minutes at room temperature(20~25°C). 4) Add 30 µL of the CD161 monoclonal antibody (10-20 µg/mL) diluted with the washingbuffer. Mix well, and incubate for 30 minutes at room temperature (20~25°C). 5) Add 1 mL of the washing buffer followed by centrifugation at 500xg for 1 minute at roomtemperature (20~25°C). Remove supernatant by careful aspiration. 6) Add 30 µL of secondary antibody (1: 40 FITC conjugated anti-mouse IgG diluted with thewashing buffer. Mix well and incubate for 15 minutes at room temperature (20~25°C). 7) Add 1 mL of the washing buffer followed by centrifugation at 500xg for 1 minute at roomtemperature (20~25°C). Remove supernatant by careful aspiration. 8) Resuspend the cells with 500 µL of the washing buffer and analyze by a flow cytometer. Protocol 2We usually use Fisher tubes or equivalents as reaction tubes for all step described below. 1) Wash the cells 3 times with washing buffer [PBS containing 2% fetal calf serum (FCS) and0. 1% NaN3]. 2) Resuspend the cells with PBS containing 25% normal goat serum, 1 mg/mL normalhuman IgG and 0. 1% NaN3 (5x10e6 cells/mL). 3) Add 20 µL of the CD161 monoclonal antibody (50 µg/mL) diluted with the washing bufferinto each tube. 4) Add 50 µL of the cell suspension into each tube. Mix well, and incubate for 30 minutesat room temperature (20~25°C). 5) Add 1 mL of the washing buffer followed by centrifugation at 500xg for 1 minute at roomtemperature (20~25°C). Remove supernatant by careful aspiration. 6) Resuspend the cells with 50 µL of the washing buffer. 7) Add 20 µL of secondary antibody (1: 10 FITC conjugated anti-mouse IgG diluted with thewashing buffer into each tube. Mix well and incubate for 15 minutes at room temperature. 8) Add 1 mL of the washing buffer followed by centrifugation at 500xg for 1 minute at roomtemperature (20~25°C). 9) Resuspend the cells with 500 µL of the washing buffer and analyze by a flow cytometer. Flow Cytometric analysis for whole blood cellsWe usually use Falcon tubes or equivalents as reaction tubes for all step described below. 1) Add 20 µL of the CD161 monoclonal antibody (50 µg/mL) diluted with washing buffer[PBS containing 2% fetal calf serum (FCS) and 0. 1% NaN3] into each tube. 2) Add 50 µL of whole blood into each tube. Mix well, and incubate for 30 minutes at roomtemperature (20~25°C). 3) Add 2 mL of the washing buffer followed by centrifugation at 500xg for 1 minute at roomtemperature (20~25°C). Remove supernatant by careful aspiration. 4) Add 50 µL of secondary antibody (1: 20 FITC conjugated anti-mouse IgG diluted with thewashing buffer. Mix well and incubate for 15 minutes at room temperature (20~25°C). 5) Lyse with OptiLyse C (for analysis on Beckman Coulter instruments) or OptiLyse B (foranalysis on BD instruments), using the procedure recommended in the respective package

Restrictions: For Research Use only

Handling

Concentration: 1.0 mg/mL

Buffer: PBS, pH 7.2 containing 50 % Glycerol without preservatives.

Preservative: Without preservative

Storage: -20 °C

Storage Comment: Store the antibody undiluted at -20 °C.
Shelf life: one year from despatch.

Expiry Date: 12 months

Target Details for CD161

Target: CD161 (KLRB1) (Killer Cell Lectin-Like Receptor Subfamily B, Member 1 (KLRB1))

Alternative Name: CD161 / KLRB1 (KLRB1 Products)

Background: CD161, also known as NKR-P1, is a receptor originally identified on the surface of natural killer cells that is also present on some subpopulations of CD8+T cells. CD161 is often used as an NK cell marker since it is present on both immature and mature NK cells. CD161 appears to mediate NK cell activation and is up-regulated by IL-12. CD161 is expressed as a disulfide homodimer of ~60 kDa while the ligand for CD161 appears to be a carbohydrate structure that includes GalNAc and GlcNAc structures. CD161+ T cells are commonly found in human liver and intestinal epithelium where they may play a role in local immunity.Synonyms: C-type lectin domain family 5 member B, CLEC5B, HNKR-P1a, Killer cell lectin-like receptor subfamily B member 1, NKRP1A, Natural killer cell surface protein P1A

Gene ID: 3820

UniProt: Q12918