The 8D7 monoclonal antibody recognizes Asialoglycoprotein receptor 1 (ASGPR 1), also known as Hepatic lectin H1 (HL-1). ASGPR 1 is an approximately 42 kDA type II integral membrane protein that is expressed on the surface of hepatic cells. It is expressed by hepatocytes on the sinusoidal-lateral plasma membrane but not on the bile canalicular membrane. ASGPR 1 plays a role in serum glycoprotein homeostasis. It functions as a subunit of the Asialoglycoprotein receptor (ASGPR) complex that binds, internalizes, and transports various glycoproteins for lysosomal degradation. The receptor may also promote hepatic infection by the binding and uptake of various viruses. The immunogen used to generate the 8D7 hybridoma was rat liver membrane extracts. Rat ASGPR consists of three polypeptide subunits (Rat hepatic lectin 1-3 (RHL1-3). The 8D7 antibody has been shown to react with a subunit-specific epitope on RHL-1. Clone 8D7 cross-reacts with human ASGPR 1.
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.