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Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD OptiBuild Brilliant reagents) 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 (Cat. No. 563794).
The T21 monoclonal antibody specifically binds to CD125 which is also known as the alpha subunit of the IL-5 Receptor (IL5RA/IL-5Rα). CD125 is a type I transmembrane protein that belongs to the hemopoietin receptor and Ig gene superfamilies. It is expressed on B cells and eosinophils. CD125 associates with the Common β chain (βc/Bc) subunit, also known as CD131, to form the functional IL-5 Receptor complex. Mouse IL-5 belongs to the hematopoietic growth factor family of cytokines and promotes the growth and differentiation of B cells and eosinophils. Upon binding, the T21 antibody blocks IL-5 binding to its receptor and can inhibit IL-5 induced B-cell proliferation and antibody production.
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.
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.