Product Information

Contents: Functional Grade Purified anti-mouse NKG2D/CD314 (activating)
Catalog Number: 16-5872
Sizes: 50 ug, 100 ug, 500 ug, 1 mg
Formulation: Phosphate buffer pH 7.2,
150 mM NaCl, No NaN₃
Storage Conditions: Store at 4°C.
Avoid repeated freeze/thaw cycles.
KEEP CONTENTS STERILE.
Endotoxin Level: Less than 0.001 ng/ug antibody, as determined by the LAL assay.
Clone: A10
Isotype: Armenian Hamster IgG

Surface staining of mouse splenocytes with anti-mouse NKG2D (A10) PE and anti-mouse pan-NK (DX5) APC. Total viable cells were used for analysis.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-5872	Affinity Purified anti-mouse NKG2D / CD314 (activating)	N/A	N/A	FA FC
16-5872	Functional Grade* Purified anti-mouse NKG2D / CD314 (activating)	N/A	N/A	FA FC
13-5872	Biotin anti-mouse NKG2D / CD314 (activating)	N/A	N/A	FC
12-5872	PE anti-mouse NKG2D / CD314 (activating)	488	575	FC
*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.

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Description

The A10 monoclonal antibody reacts with the mouse NKG2D, a lectin-like molecule expressed on both human and mouse NK cells. Mouse NKG2D binds to retinoic acid-inducible RAE-1α, -β, -γ, -δ, -ε and the minor histocompatibility molecule H60 and has the ability to costimulate multiple NK activation receptors, through the DAP12/DAP10 adaptor molecules. NKG2D is expressed by all spleen and liver NK cells, NK1.1⁺ thymocytes, in vitro activated LAK cells, and a subset of splenic NKT cells. A10 and C7 antibodies detect NK cells from all inbred strains of mice tested so far. In redirected lysis assays, A10 is reported to stimulate NK cells to kill Daudi targets. In addition, immobilized A10 is reported to induce production of GM-CSF by NK clones and LAK cells through a CD16 (FCγRIII) concomitant engagement. A10 and another hamster anti-mouse NKG2D (clone C7) compete with each other for binding to transfected cells by flow cytometric analysis, suggesting that they may bind to similar epitopes or block each other by steric hindrance. C7 (neutralizing) and A10 (activating) also exhibit different functional properties.

Expression of the NKG2D antigen on mouse peripheral NK and NKT cells can be detected by flow cytometric analysis using mAb CX5 with much brighter intensity.

Applications Reported

For research use only, not for diagnostic or therapeutic use. The A10 antibody has been reported for use in flow cytometric analysis. It has also been reported in activation in functional studies.

Applications Tested

The A10 antibody has been tested by flow cytometric analysis of mouse splenocyte suspensions. This can be used at less than or equal to 1 μg per million cells in a 100 μl total staining volume. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

References

Ho EL, Carayannopoulos LN, Poursine-Laurent J, Kinder J, Plougastel B, Smith HR, Yokoyama WM. 2002. Costimulation of multiple NK cell activation receptors by NKG2D. J Immunol. 169(7):3667-75.

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