Application

Research Sub CategoryImmunological Signaling

Anti-NK1.1 (mouse) Antibody, clone PK136, Azide Free is an antibody against NK1.1 (mouse) for use in Flow Cytometry, Activity Assay, Immunohistochemistry, Immunoprecipitation.

Research CategoryInflammation & Immunology

Activity Assay: Representative lots, when administered to mice via i.p. injection, depleted splenic natural killer (NK) cell activity in a dose-dependent manner (Xiao, T.Z., et al. (2012). Biol. Blood Marrow Transplant. 18(2):200-209; Seaman, W.E., et al. (1987). J. Immunol. 138(12):4539-4544).
Activity Assay: A representative lot activated target cell killing by cross-linking NK cell surface NK1.1 (NKR-P1C) and target cell surface Fc receptor. Ab-induced redirected lysis (AIRL) assay employing NK cells expressing both NKR-P1B and NKR-P1C prevented NK cell cytotoxicity activation by clone PK136 (Carlyle, J.R., et al. (1999). J. Immunol. 162(10):5917-5923).
Immunoprecipitation Analysis: A representative lot co-immunoprecipitated SHP-1, but not SHP-2 or SHIP, from Swiss NIH mouse-derived MNK-1 pre-NK cells upon upregulating NKR-P1B intracellular ITIM motif tyrosine phosphorylation by pervanadate treatment (Carlyle, J.R., et al. (1999). J. Immunol. 162(10):5917-5923).
Flow Cytometry Analysis: A representative lot immunostained the surface of Jurkat transfectants expressing C57BL/6J (B6) mouse-derived NKR-P1B/CD161b or Swiss NIH (Sw) mouse-derived NKR-P1C/CD161c/NK1.1, but not Jurkat transfectants expressing B6-derived NKR-P1A/CD161a (Carlyle, J.R., et al. (1999). J. Immunol. 162(10):5917-5923).
Flow Cytometry Analysis: A representative lot immunostained NKR-P1B/CD161b-expressing NK cells from Swiss NIH (Sw) mouse strain, NKR-P1C/CD161c/NK1.1-expressing NK cells from C57BL/6J (B6), as well as (B6 3 Sw)F1 cross-strain-derived NK cells expressing both NKR-P1B and NKR-P1C (Carlyle, J.R., et al. (1999). J. Immunol. 162(10):5917-5923).
Note: Clone PK136 is also available in the following conjugated forms for flow cytometry application, APC (MABF1487 & MABF1488), FITC (MABF1489 & MABF1490), PE (MABF1491), PerCP-Cy5.5 (MABF1493 & MABF1494). redFluor^® 710 (MABF1496 & MABF1497).

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

The NK cell receptor protein 1 (NKR-P1), a.k.a. CD161, molecules represent a family of type II transmembrane C-type lectin-like receptors expressed predominantly by NK cells. A total of five murine NKR-P1-coding genes (Klrb1a/b/c/d/f) have been identified, with NKR-P1A/C/F (CD161a/c/f) functioning as stimulatory receptors that possess a charged transmembrane arginine esidue thought to be important for association with the FcRγ adaptor protein, and NKR-P1B/D as two inhibitory receptors that possess a consensus cytoplasmic ITIM (L/VxYxxL/I/V) motif. The ITIM motif is shown to mediate NKR-P1B recruitment of Src homology 2 (SH2)-containing protein tyrosine phosphatase-1 (SHP-1) in a phosphorylation-dependent manner. All murine NKR-P1 proteins possess a Cys-X-Cys-Pro (CxCP) motif that mediates association with the Src-related nonreceptor protein tyrosine kinase p56lck. Both NKR-P1B and NKR-P1C functionally associate with p56lck, while mutation of the CxCP motif abolishes NKR-P1B/C-mediated signal transduction. Likewise, mutation of ITIM motif abolishes NKR-P1B SHP-1 association as well as NKR-P1B-dependent inhibitory signaling.

Immunogen

Epitope: Extracellular domain.

Mouse spleen and bone marrow cells (Koo, G.C., and Peppard, J.R. (1984). Hybridoma. 3(3):301-303).

Legal Information

GenBank is a registered trademark of United States Department of Health and Human Services

redFluor is a registered trademark of Tonbo Biotechnologies

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

Purified mouse monoclonal IgG2aκ antibody in PBS without preservatives.

Protein G purified.

Format: Purified

Quality

Evaluated by Flow Cytometry in mouse splenocytes.

Flow Cytometry Analysis: 1.0 µg of this antibody detected NK1.1-positive mouse splenocytes.

Specificity

Clone PK136 recognizes murine NKR-P1B/CD161b (UniProt P27812) and NK1.1/NKR-P1C/CD161c (UniProt P27814), but not CD161a/NKR-P1A (P27811). Upon activation by PK136 binding, NKR-P1C/NK1.1 signals to activate, while NKR-P1B signals to suppress, NK cells for cytotoxic killing of target cells (Carlyle, J.R., et al. (1999). J. Immunol. 162(10):5917-5923). Epitope is present in both spliced isoforms of murine NK1.1/CD161c reported by UniProt (P27814). Clone PK136 does not react with CD161a (by Klrb1a gene) or C57BL/6 mouse strain-derived CDC161b variant (by Klrb1b gene, also known as Klrb1d). There exists a fifth murine CD161 gene Klrb1f, reactivity of this mAb toward this gene product has not been established.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Target description

25.08 kDa (NK1.1/CD161c isoform 1) and 24.77 kDa (NK1.1/CD161c isoform 2) calculated.