CD3 Mouse anti-Human, Brilliant Ultraviolet 661, Clone: UCHT1, BD Horizon™

Description

The UCHT1 monoclonal antibody specifically binds to the human CD3 ε -chain, a 20-kDa subunit of the CD3/T cell antigen receptor complex. CD3 ε is expressed on 70-80% of normal human peripheral blood lymphocytes and 60-85% of thymocytes. Studies from the HLDA Workshop show that this antibody is mitogenic for CD3 ε -positive cells when used in conjunction with costimulatory agents such as pokeweed mitogen or anti-CD28 antibody. CD3 plays a central role in signal transduction during antigen recognition. The UCHT1 antibody stains both surface and intracellular CD3 ε unlike the other CD3 clone, HIT3a, that stains only extracellular CD3 ε .

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 630nm LP. Due to cross laser excitation of this dye, there may be significant spillover into channels detecting APC-like emissions (eg, 670/30-nm filter).

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Catalog number 563794).

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.