The 3D10 monoclonal antibody specifically binds to mouse Lymphocyte antigen 49H (Ly-49H; also known as Klra8 or Killer cell lectin-like receptor 8). The 3D10 antibody does not crossreact with related molecules such as Ly-49A, C, D or G2. Ly-49H is a type II transmembrane protein and a member of the Ly-49 C-type lectin multigene family of receptors expressed by NK cells. Cell surface Ly-49H is expressed by a subset of NK cells but not by NKT cells. Ly-49H is expressed by C57BL/6 and NWNA but not by BALB/c or DBA/2 mouse NK cells. Cell surface Ly-49H presents as a ~110 kDa disfulfide-linked homodimer and associates with signaling subunits such as DAP10 and DAP12 for optimal transduction of intracellular activation signals. Crosslinking of Ly-49H with the 3D10 antibody reportedly induces NK cell cytotoxicity and cytokine production. Ly-49H recognizes the mouse cytomegalovirus m157 glycoprotein that is expressed by infected cells and is required for protection against cytomegalovirus infection.
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.