The HIM3-4 monoclonal antibody specifically binds to CD33 which is also known as Sialic acid-binding Ig-like lectin 3 (Siglec-3), gp67, or p67. CD33 is a 67 kDa type I transmembrane glycoprotein expressed on monocytes, activated T cells, myeloid progenitors as well as mast cells. CD33 is absent on normal platelets, lymphocytes, erythrocytes and hematopoietic stem cells. This glycoprotein can reportedly function as a sialic acid-dependent cell adhesion molecule. This function can be modulated by endogenous sialoglycoconjugates when CD33 is expressed on the membrane. HIM3-4 reacts with human and monkey granulocytes and monocytes.
The antibody was conjugated to BD Horizon™ BUV737 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 737-nm. BD Horizon Brilliant BUV737 can be excited by the ultraviolet laser (355 nm) and detected with a 740/35 filter. Due to the excitation of the acceptor dye by other laser lines, there may be significant spillover into channels detecting Alexa Fluor® 700-like dyes (eg, 712/20-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 BUV737 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different BUV737 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.