The EMab-134 monoclonal antibody specifically recognizes the epidermal growth factor receptor (EGF-R). The EGF-R is a transmembrane glycoprotein of approximately 170 kDa that is expressed on most cells. It consists of a glycosylated extracellular domain which binds to epidermal growth factor (EGF), a transmembrane domain, and an intracellular domain with tyrosine-kinase activity essential for signal transduction. The epitope recognized by EMab-134 is an extracellular peptide that is not affected by glycosylation. The EGF-R plays an important role in the growth and differentiation of many cellular types. Transforming growth factor α (TGFα), vaccinia virus growth factor, and related growth factors can also bind to and signal through the EGF-R.
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.