The 3D6 monoclonal antibody specifically binds to c-Met (MET), which is also known as Hepatocyte growth factor receptor (HGFR) or Scatter factor receptor (SF receptor). c-Met is a 190 kDa single-pass type I transmembrane glycoprotein that is posttranslationally cleaved into a disulfide-linked extracellular α-chain and a transmembrane β-chain. c-Met functions as a receptor tyrosine kinase (RTK) that is normally involved in the development, regeneration, and survival of cells and tissues. c-Met is expressed on a variety of cell types including stem cells and progenitor cells, hepatocytes, keratinocytes, epithelial cells, endothelial cells, and neurons. c-Met is autophosphorylated when bound by its ligand, Hepatocyte Growth Factor (HGF). This leads to further activation of downstream signaling pathways that induce multiple responses including cellular migration, proliferation, survival, and angiogenesis. Abnormal c-Met expression or activity has been associated with tumorigenesis. The 3D6 antibody can reportedly function as an agonist by binding to and activating human but not mouse c-Met.
The antibody was conjugated to BD Horizon™ BUV737 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 737-nm. BD Horizon Brilliant BUV737 can be excited by the ultraviolet laser (355 nm) and detected with a 740/35 filter. Due to the excitation of the acceptor dye by other laser lines, there may be significant spillover into channels detecting Alexa Fluor® 700-like dyes (eg, 712/20-nm filter).
Due to spectral differences between labeled cells and beads, using BD™ CompBeads can result in incorrect spillover values when used with BD Horizon BUV737 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different BUV737 reagents (eg, CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone specific compensation controls when using these reagents.