The G9P3 monoclonal antibody specifically recognizes mouse EVA1, which is encoded by the Mpzl2 gene. EVA1 is a member of the immunoglobulin-like cell-adhesion molecule (Ig-CAM or IgSF CAM) family, members of which are anchored to the surface of epithelial and endothelial cells via a GPI domain. This class of adhesion molecules interacts intracellularly with cytoskeletal proteins and extracellularly with Ig-CAM family members, by homophilic or heterophilic binding, and with other classes of adhesion molecules such as integrins and cadherins. EVA1 is expressed on and is involved in the structural integrity of epithelia throughout the body, eg mammary glands, liver, inner ear, and the blood-brain barrier. In the thymus, EVA1 is expressed both on thymic epithelial cells (TECs) and on a subset of developing thymocytes, known as DN3. It is a low-affinity mediator of developmental signals between thymocytes and TEC that support the differentiation and survival of both developing thymocytes and the TECs. G9P3 mAb was produced in EVA1 knockout mice immunized with recombinant mouse EVA1 extracellular region, and it recognizes mouse and human EVA1 expressed on transfected cells.
The antibody was conjugated to BD Horizon™ BV711 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 711-nm. BD Horizon BV711 can be excited by the violet laser and detected in a filter used to detect Cy™5.5 / Alexa Fluor™ 700-like dyes (eg, 712/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there may be moderate spillover into the Alexa Fluor™ 700 and PerCP-Cy5.5 detectors. However, the spillover can be corrected through compensation as with any other dye combination.