The In-1 monoclonal antibody specifically binds to CD74, the MHC class II-associated Invariant chain (Ii), expressed in I-A+ cell populations. Although it is predominantly expressed intracellularly, it has also been detected on the cell surface of some B-cell lines and at low levels on a small number of B lymphocytes. In the mouse, alternatively spliced transcripts encode 31-kDa and 41-kDa isoforms, and the cell-surface CD74 has a chondroitin sulfate side chain which allows it to interact with CD44. CD74 is directly involved in the intracellular transport of MHC class II molecules and antigen presentation, and it affects the maturation of T and B lymphocytes.
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.