The 50C1 monoclonal antibody specifically binds to human CD371 which is also known as Clec12A (C-type lectin domain family 12 member A), C-type lectin-like molecule 1 (CLL-1), myeloid inhibitory C-type lectin-like receptor (MICL), or dendritic cell-associated lectin 2 (DCAL-2). It is expressed on a variety of cells including monocytes, macrophages, dendritic cells, and granulocytes and perhaps some NK cells. Clec12A is a member of the C-type lectin/C-type lectin-like domain (CTL/CTLD) superfamily. It is a 30 kDa type II transmembrane glycoprotein that has one single C-type lectin-like domain and one cytoplasmic immunoreceptor tyrosine-based inhibitory motif (ITIM). Clec12A has similarity with the β-glucan receptor (Dectin-1) and LOX-1 with high N-glycosylation. There are at least five isoforms due to alternative transcript splicing. Signaling through Clec12A can induce internalization of Clec12A, dendritic cell maturation and the production of cytokines including IL-12. Clec12A may also serve as a negative regulator of activated leukocytes recruited to sites of inflammation.
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.