Anti-NK1.1 (mouse) Antibody, clone PK136, Azide Free

Description

• 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.
• 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.
• Immunogen: Epitope: Extracellular domain.
• Application: 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).
• Quality: Evaluated by Flow Cytometry in mouse splenocytes.Flow Cytometry Analysis: 1.0 µg of this antibody detected NK1.1-positive mouse splenocytes.
• Target description: 25.08 kDa (NK1.1/CD161c isoform 1) and 24.77 kDa (NK1.1/CD161c isoform 2) calculated.
• Physical form: Format: Purified
• 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.
• Other Notes: Concentration: Please refer to lot specific datasheet.
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