Preparation And Storage
Recommended Assay Procedures
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.
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 (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Note: When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed. For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).
Product Notices
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- CF™ is a trademark of Biotium, Inc.
- BD Horizon Brilliant Ultraviolet 615 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
Companion Products
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 BUV615 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome with an Ex Max near 350 nm and an Em Max near 615 nm. BD Horizon Brilliant BUV615 can be excited by the ultraviolet laser (355 nm) and detected with a 610/20 filter and a 595 nm LP. Due to the excitation of the acceptor dye by the blue/yellow-green laser line, there may be significant spillover into channels detecting PE-CF594 like emissions (eg, 610/20-nm filter).
Development References (5)
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Adachi T, Alam R.. The mechanism of IL-5 signal transduction. Am J Physiol. 275(3 Pt 1):C623-C633. (Biology). View Reference
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Dyer KD, Garcia-Crespo KE, Percopo CM, et al. Defective eosinophil hematopoiesis ex vivo in inbred Rocky Mountain White (IRW) mice. J Leukoc Biol. 2011; 90(6):1101-1109. (Clone-specific: Flow cytometry). View Reference
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Hitoshi Y, Yamaguchi N, Mita S, et al. Distribution of IL-5 receptor-positive cells. Expression of IL-5 receptor in Ly-1 (CD5)+ B cells.. J Immunol. 1990; 144(11):4218-4225. (Immunogen: Blocking, Flow cytometry, Functional assay, Immunoprecipitation, Inhibition, Radioimmunoassay). View Reference
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Takaki S, Tominaga A, Hitoshi Y, et al. Molecular cloning and expression of the murine interleukin-5 receptor. EMBO J. 1990; 9(13):4367-4374. (Clone-specific: Cell separation, Flow cytometry). View Reference
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Wetzel, GD. Interleukin 5 regulation of peritoneal Ly-1 B lymphocyte proliferation, differentiation, and autoantibody secretion. Eur J Immunol. 1989; 19(9):1701-1707. (Biology). View Reference
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
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.