Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Mouse

Cited:

Human, Mouse, Rat, Transgenic Mouse, Xenograft

Applications

Validated:

Immunohistochemistry, Western Blot, Flow Cytometry, Agonist Activity, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, Bioassay, Functional Assay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all NKp46/NCR1 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse NKp46/NCR1
Glu22-Asn255
Accession # Q8C567

Specificity

Detects mouse NKp46/NCR1 in direct ELISAs and Western blots. In direct ELISAs, less than 15% cross-reactivity with recombinant human (rh) NKp46 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Mouse NKp46/NCR1 Antibody

Mouse NKp46/NCR1 Antibody Induces IFN-? Secretion antibody in Activated Mouse NK Cells.

Mouse NKp46/NCR1 Antibody Induces IFN-gamma Secretion in Activated Mouse NK Cells.

Mouse NKp46/NCR1 Antigen Affinity-purified Polyclonal Antibody induces IFN-? secretion in mouse natural killer (NK) cells activated with 25 ng/mL Recombinant Mouse IL-2 (Catalog # 402-ML) and 25 ng/mL Recombinant Mouse IL-12 (Catalog # 419-ML), in a dose-dependent manner, as measured using the Quantikine Mouse IFN-? ELISA Kit (Catalog # MIF00). The ED50 for this effect is typically 0.4-2.4 µg/mL.
Detection of NKp46/NCR1 antibody in Mouse DX5/CD49b+Splenocytes antibody by Flow Cytometry.

Detection of NKp46/NCR1 in Mouse DX5/CD49b+Splenocytes by Flow Cytometry.

Mouse DX5/CD49b+splenocytes were stained with Goat Anti-Mouse NKp46/NCR1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2225, filled histogram) or control antibody (Catalog # AB-108-C, open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108).
NKp46/NCR1 antibody in Mouse Spleen by Immunohistochemistry (IHC-Fr).

NKp46/NCR1 in Mouse Spleen.

NKp46/NCR1 was detected in perfusion fixed frozen sections of mouse spleen using Goat Anti-Mouse NKp46/NCR1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2225) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Detection of Mouse NKp46/NCR1 by Immunocytochemistry/Immunofluorescence

Detection of Mouse NKp46/NCR1 by Immunocytochemistry/Immunofluorescence

Acutely Cultured Embryonic but Not Adult DRG Neurons Reveal Susceptibility to NK-Mediated Cytotoxicity by RAE1(A) Immunolabeling of co-culture (4 h) between embryonic (top) or adult (bottom) DRG neurons ( beta -tubulin, magenta) and either freshly isolated (control) or IL-2-stimulated natural killer (NK) cells (NKp46, green). The inset shows a high-magnification image of NK cell in contact with embryonic DRG neurite.(B) LDH-release cytotoxicity assay of acutely cultured (1 day in vitro) embryonic (top) and adult (bottom) DRG at various Effector (NK):Target (DRG) (E:T) ratios. Matched two-way ANOVA: embryonic DRG, F(1,10) = 100.01, p < 0.0001); adult DRG, F(1,10) = 1.25, p = 0.2982). Three replicate co-cultures for each DRG group.(C) Still images of in vitro time-lapse confocal Ca2+ imaging of rhodamine 3 AM-loaded embryonic (top) and adult (bottom) DRG (magenta) co-cultured with IL-2-stimulated NK cells (green) isolated from adult male NKp46-YFP mice.(D) Frequency histogram (30 s time bins) of neurite Ca2+ events in embryonic (top) and adult (bottom) DRG during NK co-culture. Cumulative area under the curve (right). Student’s paired t test; t = 2.290, p = 0.045. n = 6 fields of view from two repeat co-cultures per group.(E) RT-PCR of mRNA transcripts in freshly isolated splenic NK cells and embryonic and adult DRG.(F) qRT-PCR shows higher Raet1 mRNA expression in embryonic compared to adult DRG tissue. Student’s paired t test; t = 16.16, p < 0.0001. n = 5 mice, or replicates per group.(G) Western blot of embryonic and adult mouse DRG tissue (40 μg loading) with pan-RAE1 antibody and beta -actin control. Images are representative of three independent experiments.(H) Selective siRNA knockdown reduces RAE1 protein (top) and Raet1 mRNA (bottom) expression in embryonic DRG (2 d culture). Student’s unpaired t test; t = 9.060, p = 0.0008. n = 3 mice, or replicates per group.(I) LDH-release cytotoxicity assay of negative control or Raet1-selective siRNA knockdown embryonic DRG. Three replicate co-cultures for each siRNA group. Matched two-way ANOVA F(1,10) = 133.85, p < 0.0001).See also Figure S1. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30712871), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of NKp46/NCR1 by Immunohistochemistry

Detection of NKp46/NCR1 by Immunohistochemistry

mNemvaleukin alone and combination treatments expand tumor-infiltrating NK cells in SCLC. (A) Schematic showing experimental design for immune profiling. After confirming tumor burden by MRI, mice were randomized into vehicle, chemotherapy, mNemvaleukin, and combo (chemotherapy+mNemvaleukin) treatment groups. Tumor nodules were collected and tumor-infiltrating lymphocytes were analyzed by flow cytometry at day 7 after treatment initiation. (B) Representative plots of infiltrating NK cells (CD3−CD49b+) are shown on indicated treatment. (C, D) Frequencies of total infiltrating NK cells (C), activated NK cells (D) and expression of Ki-67+ (E) in NK cells are presented. Two independent cohorts were performed (n=4 in total for each group). (F) IHC staining of NKp46 on tumor samples treated with vehicle, chemotherapy, mNemvaleukin, and combo regimens. Data shown as means±SEM. *P<0.05, **P<0.01, ***P<0.001. IHC, Immunohistochemistry; mNemvaleukin, mouse version of nemvaleukin; NK, natural killer; ns, not significant (unpaired two-tailed t test); SCLC, small cell lung cancer. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36472839), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of NKp46/NCR1 by Immunohistochemistry

Detection of NKp46/NCR1 by Immunohistochemistry

mNemvaleukin alone and combination treatments expand tumor-infiltrating NK cells in SCLC. (A) Schematic showing experimental design for immune profiling. After confirming tumor burden by MRI, mice were randomized into vehicle, chemotherapy, mNemvaleukin, and combo (chemotherapy+mNemvaleukin) treatment groups. Tumor nodules were collected and tumor-infiltrating lymphocytes were analyzed by flow cytometry at day 7 after treatment initiation. (B) Representative plots of infiltrating NK cells (CD3−CD49b+) are shown on indicated treatment. (C, D) Frequencies of total infiltrating NK cells (C), activated NK cells (D) and expression of Ki-67+ (E) in NK cells are presented. Two independent cohorts were performed (n=4 in total for each group). (F) IHC staining of NKp46 on tumor samples treated with vehicle, chemotherapy, mNemvaleukin, and combo regimens. Data shown as means±SEM. *P<0.05, **P<0.01, ***P<0.001. IHC, Immunohistochemistry; mNemvaleukin, mouse version of nemvaleukin; NK, natural killer; ns, not significant (unpaired two-tailed t test); SCLC, small cell lung cancer. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36472839), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse NKp46/NCR1 Antibody

Application
Recommended Usage

Agonist Activity

0.4-2.4 µg/mL
Sample: Mouse natural killer (NK) cells activated with Recombinant Mouse IL-2 (Catalog # 402-ML) and Recombinant Mouse IL-12 (Catalog # 419-ML)

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

2.5 µg/106 cells
Sample: Mouse DX5/CD49b+ splenocytes

Immunohistochemistry

3-15 µg/mL
Sample: Perfusion fixed frozen sections of mouse spleen

Western Blot

0.1 µg/mL
Sample: Recombinant Mouse NKp46/NCR1 Fc Chimera (Catalog # 2225-NK)

Reviewed Applications

Read 2 reviews rated 5 using AF2225 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & 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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: NKp46/NCR1

NKp46, along with NKp30 and NKp44, are activating receptors that have been collectively termed the natural cytotoxicity receptors (NCR) (1). These receptors are expressed almost exclusively by NK cells and play a major role in triggering some of the key lytic activities of NK cells. In human systems, the CD56dimCD16+ subpopulation that makes up the majority of NK cells in the peripheral blood and spleen expresses NKp46 in both resting and activated states (2). The main NK cell population of the lymph node (CD56brightCD16-) expresses low levels of NKp46 in resting cells, but expression is upregulated by IL-2. Mouse NKp46, also known as MAR-1 (3), is a type I transmembrane protein with two extracellular Ig-like domains. It has a positive charge in its transmembrane domain that permits association with the ITAM-bearing signal adapter proteins, CD3 zeta and Fc epsilon RI gamma (4). Studies with neutralizing antibodies indicate that the three NCR are primarily responsible for triggering the NK-mediated lysis of many human tumor cell lines. Blocking any of the NCRs individually resulted in partial inhibition of tumor cell lysis, but nearly complete inhibition of lysis was observed if all three receptors were blocked simultaneously (5). NKp46 has also been implicated in recognition of virus-infected cells through its capacity to bind to viral hemagglutinins (6-8).

References

  1. Moretta, L. and A. Moretta (2004) EMBO J. 23:255.
  2. Ferlazzo, G. et al. (2004) J. Immunol. 172:1455.
  3. Biassoni, R. et al. (1999) Eur. J. Immunol. 29:1014.
  4. Westgaard, I. et al. (2004) J. Leukoc. Biol. PMID 15356098.
  5. Pende, D. et al. (1999) J. Exp. Med. 190:1505.
  6. Arnon, T. et al. (2004) Blood 103:664.
  7. Arnon, T. et al. (2001) Eur. J. Immunol. 31:2680.
  8. Mandelboim, O. et al. (2001) Nature 409:1055.

Alternate Names

CD335, Ly94, MAR-1, NCR1

Entrez Gene IDs

9437 (Human); 17086 (Mouse); 100141419 (Porcine); 102115479 (Cynomolgus Monkey)

Gene Symbol

NCR1

UniProt

Q8C567

Additional NKp46/NCR1 Products

Product Documents for Mouse NKp46/NCR1 Antibody

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Product Specific Notices for Mouse NKp46/NCR1 Antibody

For research use only

Citations for Mouse NKp46/NCR1 Antibody

Customer Reviews for Mouse NKp46/NCR1 Antibody (2)

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  • Mouse NKp46/NCR1 Antibody
    Name: Efstathios Stamatiades
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Salivary gland tissue
    Species: Mouse
    Verified Customer | Posted 07/10/2019
    PLP-fixed salivary gland cryosections were stained with anti NKp46/NCR1 for 4h at room temperature. After washing, the samples were stained with Donkey anti-goat alexa546 2ndary antibody for 1h at room temperature. NKp46 in red, CD3 in green &amp; Hoechst in blue.
    Mouse NKp46/NCR1 Antibody AF2225
  • Name: Anonymous
    Application: Immunohistochemistry-Frozen
    Sample Tested: See PMID 23902341
    Species: Mouse
    Verified Customer | Posted 01/06/2015

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