Staining of mouse splenocytes with Anti-Mouse CD49b (Integrin α2) APC (cat. 17-5971) and 0.25 µg of Anti-Mouse CD314 (NKG2D) PE. Total viable cells were used for analysis.
Contents: Anti-Mouse CD314 (NKG2D) PE Catalog Number: 12-5872 Concentration: 0.2 mg/ml Formulation: Phosphate buffer pH 7.2, 150 mM NaCl, 0.09% NaN3 Storage Conditions: Store at 2-8°C. Do not freeze. Light sensitive material. Clone: A10 Host/Isotype: Armenian Hamster IgG
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12-5872-81
50 ug
12-5872-82
100 ug
12-5872-83
200 ug
Note: Several countries will continue to be supplied via distributors. Country specific prices may apply.
*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 Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.
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.
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 0.5 μg per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 105 to 108 cells/test. 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.
