CD314 (NKG2D) Monoclonal Antibody (A10), Functional Grade, eBioscience™, Invitrogen™

Description

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-1 alpha, beta, gamma, delta, epsilon 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 (FC gamma RIII) 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: 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 splenocytes. This can be used at less than or equal to 1 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 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Endotoxin: Less than 0.001 ng/ug antibody as determined by the LAL assay. Storage and handling: Use in a sterile environment. Filtration: 0.2 μm post-manufacturing filtered. Purity: Greater than 90%, as determined by SDS-PAGE. Endotoxin Level: Less than 0.001 ng/μg antibody, as determined by LAL assay. Aggregation: Less than 10%, as determined by HPLC. Natural killer (NK) cells are lymphocytes that can mediate lysis of certain tumor cells and virus-infected cells without previous activation. They can also regulate specific humoral and cell-mediated immunity. NK cells preferentially express several calcium-dependent (C-type) lectins, which have been implicated in the regulation of NK cell function. This gene encodes a member of the NKG2 family, and the encoded transmembrane protein is characterized by a type II membrane orientation (extracellular C terminus) and the presence of a C-type lectin domain. The NKG2 gene family is located within the NK complex, a region that contains several C-type lectin genes preferentially expressed in NK cells.