The MECA-367 monoclonal antibody specifically recognizes mucosal vascular addressin MAdCAM-1. In the fetus and neonate, MAdCAM-1 is the predominant vascular addressin on the high endothelial venules (HEV) of peripheral lymph nodes. In adult mice, MAdCAM-1 is preferentially expressed in mucosal lymphoid tissues and lamina propria; it is also expressed on sinus-lining cells in the spleen. MAdCAM-1 expression is upregulated on the HEV of peripheral lymph nodes in adult NOD mice4 and is involved in the development of diabetes and insulitis. Furthermore, there is evidence that IFN-γ can induce MAdCAM-1 expression in non-mucosal sites in adult mice. MAdCAM-1 is a predominant ligand for integrin α4β7, a lymphocyte mucosal homing receptor, and a facultative ligand for CD62L (L-selectin). MECA-367 mAb binds to the first domain of MAdCAM-1 and blocks MAdCAM-1-dependent binding in vitro and lymphocyte homing to Peyer's patch HEV in vivo. Source of the immunogen was endothelial cells from BALB/c mouse mesenteric and peripheral lymph nodes.
The antibody was conjugated to BD Horizon™ BUV563 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 which has an Ex Max of 348 nm and an acceptor dye. The tandem has an Em Max at 563 nm. BD Horizon BUV563 can be excited by the 355 nm ultraviolet laser. On instruments with a 561 nm Yellow-Green laser, the recommended bandpass filter is 585/15 nm with a 535 nm long pass to minimize laser light leakage. When BD Horizon BUV563 is used with an instrument that does not have a 561 nm laser, a 560/40 nm filter with a 535 nm long pass may be more optimal. Due to the excitation and emission characteristics of the acceptor dye, there may be spillover into the PE and PE-CF594 detectors. However, the spillover can be corrected through compensation as with any other dye combination.