The Neu 24.7 monoclonal antibody specifically recognizes HER-2/neu, which is also known as p185HER2 or CD340. HER-2/neu is a ~185 kDa type I transmembrane glycoprotein that is encoded by ERBB2 (erb-b2 receptor tyrosine kinase 2) which belongs to the ERBB receptor kinase family. The HER-2/neu antigen is expressed on some breast cancer cells. Amplification and/or over expression of HER-2/neu occurs in multiple human malignancies. Overexpression is reported to be strongly correlated with progression of human ovarian and breast carcinomas. Anti-HER-2/neu stains the external domain of this protein on cells such as the strongly expressing line SKBR3 and the weakly expressing line MCF7. HER-2/neu is also expressed on normal bone marrow mesenchymal stem cells (MSC).
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.