The 371805 monoclonal antibody specifically recognizes Ephrin type-A receptor 2 (EphA2). EphA2 is a ~130 kDa type I transmembrane glycoprotein that is encoded by EPHA2 (EPH receptor A2) which belongs to ephrin receptor subfamily within the receptor tyrosine kinase (RTK) family. EphA2 is also known as Epithelial cell kinase (ECK) or Epithelial cell receptor protein tyrosine kinase. The extracellular region of EphA2 contains an N-terminal globular domain, a cysteine-rich domain and 2 fibronectin type III repeats, whereas the cytoplasmic domain contains two tyrosine residues adjacent to the membrane that function as major autophosphorylation sites and a kinase domain. EphA2 is variably expressed by epithelial cells, dendritic cells, Langerhans cells, keratinocytes, and endothelial cells. EphA2 functions as a receptor for glycophosphatidylinositol (GPI) membrane-linked members of the Ephrin-A family, including Ephrins A1-A5. EphA2 is involved in regulating cellular growth, adhesion, migration, survival, and plays a role in angiogenesis. Its expression may be upregulated on vascular endothelium in certain breast, prostate, and colon cancers as well as on some metastatic tumor cells.
The antibody was conjugated to BD Horizon™ BUV661 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 661-nm. BD Horizon Brilliant BUV661 can be excited by the ultraviolet laser (355 nm) and detected with a 670/25 filter and a 630 nm LP. Due to cross laser excitation of this dye, there may be significant spillover into channels detecting APC-like emissions (eg, 670/25-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 BUV661 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different BUV661 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.