NKp30/NCR3 Antibody (210845) [Unconjugated]

Human NKp30 Monoclonal Antibody induces IFN‑ gamma secretion in the NK-92 human natural killer lymphoma cell line in a dose-dependent manner, as measured using the Quantikine Human IFN‑ gamma ELISA Kit (Catalog # DIF50). The ED50 for this effect is typically 0.2‑1.2 μg/mL.

Human peripheral blood lymphocytes were stained with either (A) Mouse Anti-Human NKp30/NCR3 Monoclonal Antibody (Catalog # MAB1849) or (B) Mouse IgG2A Isotype Control (MAB003) followed by anti-Mouse IgG APC-conjugated secondary antibody (F0101B) and Mouse Anti-Human NCAM-1/CD56 PE-conjugated Monoclonal Antibody (FAB2408P). Staining was performed using our Staining Membrane-associated Proteins protocol.

beta -1,3-glucan is required for NKp30-mediated killing of Cryptococcus. g NKp30 binding to C. neoformans vs. C. albicans analyzed using flow cytometry. The experiment was performed twice. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29467448), licensed under a CC-BY license. Not internally tested by R&D Systems.

beta -1,3-glucan is required for NKp30-mediated killing of Cryptococcus. f Anti-NKp30 mAb (1C01) blocked NKp30 binding to beta -1,3-GB. Recombinant NKp30 was incubated with 1C01 before being applied to beads conjugated with beta -1,3-glucan. The presence of NKp30 on beta -1,3-GB was detected using the polyclonal anti-NKp30 antibody. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29467448), licensed under a CC-BY license. Not internally tested by R&D Systems.

beta -1,3-glucan increases expression & clustering of NKp30 in NK . a NKp30 expression in YT in response to beta -1,3-glucan. YT treated with beta -1,3-glucan (III & IV) & expression of NKp30 using 1C01 was compared to untreated (I), or untreated labeled with isotype control (II). b WB analysis of NKp30 expression in YT in response to beta -1,3-glucan. YT incubated with or without beta -1,3-glucan for various times. The experiment was repeated three times with similar results. c NKp30 labeling at the synapse between a representative single YT cell & a single beta -1,3-glucan-conjugated bead. 1C01 was used to label NKp30. The location of the bead is shown with a white arrow. Bar = 5 µm. d Proximity of NKp30 expression in relation to the radius defined by the center of the IS (n = 21 ) in panel c. The method for determining the angle is illustrated in Supplementary Fig. 6C. Clustering was defined when the peak of fluorescence was at an angle <45° (blue bars in the left). e Frequency of NKp30 clustering of all YT cell conjugates with beta -1,3-glucan-conjugated or unconjugated beads as analyzed in panel c, as defined in panel d. f TIRF microscopic analysis of NKp30 expression at the interface between YT & beta -1,3-glucan or control. YT loaded into a glass chamber that had been coated with beta -1,3-glucan or mannan incubated at 37 °C for 30 min, fixed & stained for NKp30 using 1C01. Bar = 10 µm. g Fluorescent intensity of NKp30 from panel f in arbitrary units. Distance is defined in Supplementary Fig. 6A-B. All experiments repeated three times with similar results. TIRF total internal reflection fluorescence, poly-l-lysine coating alone or mannan coating served as control, Au arbitrary unit, Arrow bead, SD spinning disc microscopy, DIC differential interference contrast digital image, IS immunological synapse Image collected & cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29467448), licensed under a CC-BY license. Not internally tested by R&D Systems.

beta -1,3-glucan is required for NKp30-mediated killing of Cryptococcus. e Flow cytometric analysis of a recombinant NKp30-Fc fusion protein binding to beta -glucan-conjugated beads ( beta -1,3-GB) compared to unconjugated polystyrene beads as control. NKp30-Fc on the beads was detected with anti-NKp30 antibody (1C01). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29467448), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: Flow, CyTOF-ready, Func
• Clone: 210845
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

NKp30/NCR3 Antibody (210845) [Unconjugated] Summary

• Additional Information: Clone 210845 was used by HLDA to establish CD designation
• Immunogen: Mouse T cell hybridoma transfected with human NKp30/NCR3 and Mouse myeloma cell line NS0-derived recombinant human NKp30 Fc Chimera
• Specificity: Detects human NKp30/NCR3 on cell transfectants and peripheral blood NK cells.
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Gene: NCR3
• Purity Statement: Protein A or G purified from hybridoma culture supernatant
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Agonist Activity 0.2-1.2 ug/mL
  • CyTOF-ready
  • Flow Cytometry 0.25 ug/10^6 cells

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for NKp30/NCR3 Antibody (210845) [Unconjugated]

• Activating natural killer receptor p30
• CD337 antigen
• CD337
• CD337activating NK-A1 receptor
• LY117,1C7NK-p30
• lymphocyte antigen 117
• natural cytotoxicity triggering receptor 3
• Natural killer cell p30-related protein
• NCR3
• NKp30
• NKp30MALS

Background

NKp30, along with NKp44 and NKp46, constitute a group of receptors termed “Natural Cytotoxicity Receptors” (1). These receptors play a major role in triggering NK‑mediated killing of most tumor cells lines. NKp30 is a type I transmembrane protein having a single extracellular V-like immunoglobulin domain (2). A physical association with the ITAM-bearing accessory protein, CD3 zeta , occurs via a charged residue in the NKp30 transmembrane domain. Ligation of NKp30 with a specific antibody results in phosphorylation of CD3 zeta (3). NKp30 is expressed on both resting and activated NK cells of the CD56^dim, CD16⁺ subset that account for more that 85% of NK cells found in peripheral blood and spleen (4). NKp30 is absent from the CD56^bright, CD16^- subset that constitutes the majority of NK cells in lymph node and tonsil, however, its expression is up-regulated in these cells upon IL-2 activation (4). Studies with neutralizing antibodies reveal that NKp30 is partially responsible for triggering lytic activity against several tumor cell types and that it is the main receptor responsible for NK‑mediated lysis of immature dendritic cells (2, 5). The ligand(s) recognized by NKp30 has not been described.

1. Moretta, L. and A. Moretta (2004) EMBO J. 23:255.
2. Pende, D. et al. (1999) J. Exp. Med. 190:1505.
3. Augugliaro, R. et al. (2003) Eur. J. Immunol. 33:1235.
4. Ferlazzo, G. et al. (2004) J. Immunol. 172:1455.
5. Ferlazzo, G. et al. (2002) J. Exp. Med. 195:343.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for NKp30/NCR3 Antibody (MAB1849)(8)

Publications using MAB1849

• R Chimienti, T Baccega, S Torchio, F Manenti, S Pellegrini, A Cospito, A Amabile, MT Lombardo, P Monti, V Sordi, A Lombardo, M Malnati, L Piemonti Engineering of immune checkpoints B7-H3 and CD155 enhances immune compatibility of MHC-I-/- iPSCs for beta cell replacement Cell Reports, 2022-09-27;40(13):111423. 2022-09-27 [PMID: 36170817] (ICC, Human)
• N Kim, MY Kim, YU Cho, W Chen, KH Lee, HS Kim PVR and ICAM-1 on Blast Crisis CML Stem and Progenitor Cells with TKI Resistance Confer Susceptibility to NK Cells Cancers (Basel), 2020-07-16;12(7):. 2020-07-16 [PMID: 32708713] (Flow Cytometry, Human)
• Z Djaoud, LA Guethlein, A Horowitz, T Azzi, N Nemat-Gorg, D Olive, D Nadal, PJ Norman, C Münz, P Parham Two alternate strategies for innate immunity to Epstein-Barr virus: One using NK cells and the other NK cells and ?? T cells J. Exp. Med., 2017-05-03;0(0):. 2017-05-03 [PMID: 28468758] (Neutralization, Human)
• Maryam Salimi, Luzheng Xue, Helen Jolin, Clare Hardman, David J. Cousins, Andrew N. J. McKenzie et al. Group 2 innate lymphoid cells express functional NKp30 receptor inducing type 2 cytokine production1 The Journal of Immunology 2016-01-01 [PMID: 26582946] (Flow Cytometry, Human)
• Herrmann , Julia, Berberich , Hannah, Hartmann , Jessica, Beyer , Steffen, Davies , Karen, Koch , Joachim Homo-oligomerization of the activating natural killer cell receptor NKp30 ectodomain increases its binding affinity for cellular ligands. J Biol Chem, 2013-11-25;289(2):765-77. 2013-11-25 [PMID: 24275655] (ELISA Development, Flow Cytometry, IHC, IHC-Fr, Human, Insect, Moth - Trichoplusia ni (Cabbage Looper), Mouse)
• Shi J, Tricot GJ, Garg TK, Malaviarachchi PA, Szmania SM, Kellum RE, Storrie B, Mulder A, Shaughnessy JD, Barlogie B, van Rhee F Bortezomib down-regulates the cell-surface expression of HLA class I and enhances natural killer cell-mediated lysis of myeloma. Blood, 2007-10-18;111(3):1309-17. 2007-10-18 [PMID: 17947507] (Neutralization, Human)
• Warren HS, Warren</LastName><ForeNam HS</Initia, Bettadapura J A novel binding assay to assess specificity of monoclonal antibodies. J. Immunol. Methods, 2005-08-15;305(1):33-8. 2005-08-15 [PMID: 16125193] (Epitope Mapping, Human)
• Warren HS, Jones AL, Freeman C, Bettadapura J, Parish CR Evidence that the cellular ligand for the human NK cell activation receptor NKp30 is not a heparan sulfate glycosaminoglycan. J. Immunol., 2005-07-01;175(1):207-12. 2005-07-01 [PMID: 15972650] (Flow Cytometry, Human)

Bioinformatics

• Gene Symbol: NCR3