The miap301 monoclonal antibody specifically binds to the extracellular domain of CD47, also known as Integrin-Associated Protein (IAP). CD47 is a 50-kDa plasma membrane protein with an extracellular immunoglobulin variable region-like domain, a multiple membrane-spanning segment, and a short C-terminal cytoplasmic tail. IAP is often physically associated with β3 integrins, and it may act as a transducer element in activation mediated by these integrins. CD47 is expressed by a wide variety of cell types, including some cells lacking integrins, such as erythrocytes. The cytoplasmic tail region is expressed as alternatively spliced forms, which are differentially expressed in diverse tissues, suggesting that IAP may have distinct functions in various tissues. CD47 and its ligand, CD172a, are proposed to mediate bi-directional signaling to modify neural synaptic activity and regulate the phagocytic activities of macrophages.
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.