Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Human, Mouse, Transgenic Mouse

Applications

Validated:

Western Blot, Blockade of Receptor-ligand Interaction, Flow Cytometry, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Frozen, Western Blot, Neutralization, Bioassay, Functional Assay

Label

Unconjugated

Antibody Source

Monoclonal Rat IgG2B Clone # 191004
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Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse NKG2D
Phe94-Val232
Accession # O54709

Specificity

Detects mouse NKG2D in direct ELISAs and Western blots. Does not cross-react with recombinant human NKG2D.

Clonality

Monoclonal

Host

Rat

Isotype

IgG2B

Endotoxin Level

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

Scientific Data Images for Mouse NKG2D/CD314 Antibody

Detection of NKG2D/CD314 by Flow Cytometry

Detection of NKG2D/CD314 by Flow Cytometry

NK Require NKG2D to Control Early Virus Dissemination & for Optimal Cytotoxicity but Not Activation(A) B6 mice infected with 3,000 pfu ECTV. On day 5 PI, mice pulsed with BrdU for 3 h & their spleens analyzed by flow cytometry. Plots are gated on CD3ɛ− NK1.1+. Data correspond to pools of three mice from three individual experiments.(B) Increased viral titers of infected mice with NKG2D blockade in vivo. Intact B6 mice, B6 mice with NKG2D blockade, B6 mice depleted of NK, B6 mice depleted of T (treated with anti-CD4 + anti-CD8 mAbs), or depleted of T & with NKG2D blockade infected with 3,000 pfu ECTV & the viral titers in spleen determined on day 3 PI. Data are the average ± SD of three individual mice & are representative of two similar experiments.(C) The indicated mice infected with 3,000 pfu ECTV, & the NK responses in the D-LN determined at 2 d PI. Upper panel: Dot plot showing the proportion of NK (DX5+, CD3e−) in the D-LN of infected & control uninfected mice. Lower panel: GzB & IFN-gamma production by NK. correspond to the DX5+, CD3ɛ− gate of the upper panels. Data correspond to pools of three mice & are representative of three similar experiments.(D) NK purified from spleens (5 d PI) of ECTV-infected intact or NKG2D-blocked mice & stained as indicated. Filled histogram: isotype-matched control Ig; gray line: NKG2D-blocked mice; black line: intact mice.(E) NK purified from spleens of intact or NKG2D-blocked mice as indicated & used as effectors in a 4-h 51Cr release assay against the indicated targets. Data correspond to pools of three mice & are representative of three experiments.(F) As in (D), but the NK purified from untreated mice & a neutralizing anti-NKG2D mAb was added to the in vitro cytotoxicity assays, as indicated. Image collected & cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/18266471), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of NKG2D/CD314 by Flow Cytometry

Detection of NKG2D/CD314 by Flow Cytometry

NK Require NKG2D to Control Early Virus Dissemination & for Optimal Cytotoxicity but Not Activation(A) B6 mice infected with 3,000 pfu ECTV. On day 5 PI, mice pulsed with BrdU for 3 h & their spleens analyzed by flow cytometry. Plots are gated on CD3ɛ− NK1.1+. Data correspond to pools of three mice from three individual experiments.(B) Increased viral titers of infected mice with NKG2D blockade in vivo. Intact B6 mice, B6 mice with NKG2D blockade, B6 mice depleted of NK, B6 mice depleted of T (treated with anti-CD4 + anti-CD8 mAbs), or depleted of T & with NKG2D blockade infected with 3,000 pfu ECTV & the viral titers in spleen determined on day 3 PI. Data are the average ± SD of three individual mice & are representative of two similar experiments.(C) The indicated mice infected with 3,000 pfu ECTV, & the NK responses in the D-LN determined at 2 d PI. Upper panel: Dot plot showing the proportion of NK (DX5+, CD3e−) in the D-LN of infected & control uninfected mice. Lower panel: GzB & IFN-gamma production by NK. correspond to the DX5+, CD3ɛ− gate of the upper panels. Data correspond to pools of three mice & are representative of three similar experiments.(D) NK purified from spleens (5 d PI) of ECTV-infected intact or NKG2D-blocked mice & stained as indicated. Filled histogram: isotype-matched control Ig; gray line: NKG2D-blocked mice; black line: intact mice.(E) NK purified from spleens of intact or NKG2D-blocked mice as indicated & used as effectors in a 4-h 51Cr release assay against the indicated targets. Data correspond to pools of three mice & are representative of three experiments.(F) As in (D), but the NK purified from untreated mice & a neutralizing anti-NKG2D mAb was added to the in vitro cytotoxicity assays, as indicated. Image collected & cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/18266471), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse NKG2D/CD314 Antibody

Application
Recommended Usage

Blockade of Receptor-ligand Interaction

In a functional ELISA, 0.05-0.15 µg/mL of this antibody will block 50% of the binding of 100 ng/mL of biotinylated Recombinant Mouse Rae-1 gamma Fc Chimera to immobilized Recombinant Mouse NKG2D Fc Chimera (Catalog # 139‑NK) coated at 4 µg/mL (100 µL/well). At 2 μg/mL, this antibody will block >90% of the binding.

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: NK1.1+ mouse splenic NK cells

Western Blot

1 µg/mL
Sample: Recombinant Mouse NKG2D/CD314 Fc Chimera (Catalog # 139-NK)
under non-reducing conditions only

Reviewed Applications

Read 1 review rated 5 using MAB1547 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.


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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: NKG2D/CD314

NKG2D, also known as CD314, is a type II transmembrane protein with an extracellular C-type lectin-like domain. It occurs as a disulfide-linked homodimer that associates with the transmembrane DAP10 (DNAX-activator protein 10) adapter protein to deliver an activating signal. This protein shares approximately 25% amino acid sequence identity with a number of other type II lectin-like proteins that are encoded by genes within the natural killer complex on mouse chromosome 6. NKG2D is expressed on NK cells, where it functions as an activating receptor to trigger cytolytic activity and cytokine secretion, and on some T cell subsets, where it acts as a co‑stimulatory receptor complementing T cell receptor signaling. Several ligands have now been identified for mouse NKG2D including H60 and Rae 1 alpha, beta, gamma, delta, and epsilon. All of these ligands are cell-surface proteins distantly related to MHC class I. However, they do not bind peptide or associate with beta 2-microglobulin. Ligand expression is up-regulated in many transformed cell lines and also during conditions of stress such as heat shock or viral infection. In vivo, tumor models demonstrate that NKG2D functions in anti-tumor surveillance (1-5).

References

  1. Bauer, S. et al. (1999) Science 285:727.
  2. Wu, J. et al. (1999) Science 285:730.
  3. Diefenbach, A. et al. (2001) Nature 413:165.
  4. Vivier, E. et al. (2002) Curr. Opin. Immunol. 14:306.
  5. NKG2D and its Ligands; www.RnDSystems.com.

Long Name

Natural Killer G2D

Alternate Names

CD314, D12S2489E, KLRK1

Entrez Gene IDs

22914 (Human); 27007 (Mouse); 102120479 (Cynomolgus Monkey)

Gene Symbol

KLRK1

UniProt

Additional NKG2D/CD314 Products

Product Documents for Mouse NKG2D/CD314 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Mouse NKG2D/CD314 Antibody

For research use only

Citations for Mouse NKG2D/CD314 Antibody

Customer Reviews for Mouse NKG2D/CD314 Antibody (1)

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  • Mouse NKG2D/CD314 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Dendritic epidermal T cells
    Species: Mouse
    Verified Customer | Posted 10/24/2021
    Mouse NKG2D/CD314 Antibody MAB1547

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