The RUU-PL7F12 monoclonal antibody specifically recognizes CD61, a 110 kDa type I transmembrane glycoprotein, also known as Glycoprotein IIIa (gpIIIa), the common β-subunit (integrin β3-chain) of the gpIIb/IIIa complex and the vitronectin receptor (VNR). The gpIIb/IIIa complex and the VNR are integrins, ie, α/β-heterodimeric glycoprotein complexes that are involved in cell adhesion. With the CD41 antigen (gpIIb or αIIb), the CD61 antigen forms the gpIIb/IIIa complex, which acts as a receptor for fibrinogen, von Willebrand factor (vWf), fibronectin, and vitronectin on activated platelets. With the CD51 antigen (VNR α-chain or αv), the CD61 antigen forms the VNR, which mediates activation-independent cell adhesion to vitronectin, vWf, fibrinogen, and thrombospondin. The CD61 antigen is found on all normal resting and activated platelets. Platelets from individuals with Glanzmann's thrombasthenia show a >90% reduction of binding of CD61, and heterozygote carriers of the disorder show approximately 50% reduction. The CD61 antigen is also found on endothelial cells, megakaryocytes, and on some myeloid, erythroid, and T-lymphoid leukemic cell lines.
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.