Preparation And Storage
Recommended Assay Procedures
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).
Product Notices
- This antibody was developed for use in flow cytometry.
- 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.
Companion Products
The 20/70 monoclonal antibody specifically binds to CD88, which is also known as, Complement component 5a receptor 1 (C5ar1), C5a anaphylatoxin chemotactic receptor, or C5a receptor (C5aR). CD88 belongs to the rhodopsin family of seven-transmembrane G protein-coupled receptors and mediates its effects by binding to the complement activation product, C5a. CD88 is expressed on leucocytes including, neutrophils, monocytes, macrophages, and eosinophils. CD88 may also be expressed on alveolar epithelial cells, endothelial cells, neural stem cells, oligodendrocytes, and on parenchymal cells of lung, liver, kidney and heart. Binding of the proinflammatory C5a anaphylatoxin to CD88 results in G-protein coupling and intracellular signal transduction that mediates intracellular calcium release, chemotaxis, degranulation, cytokine production, and superoxide anion production. The 20/70 antibody can block binding of C5a to CD88 and thus inhibit its biological functions.
The antibody was conjugated to BD Horizon™ BUV563 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 which has an Ex Max of 348 nm and an acceptor dye. The tandem has an Em Max at 563 nm. BD Horizon BUV563 can be excited by the 355 nm ultraviolet laser. On instruments with a 561 nm Yellow-Green laser, the recommended bandpass filter is 585/15 nm with a 535 nm long pass to minimize laser light leakage. When BD Horizon BUV563 is used with an instrument that does not have a 561 nm laser, a 560/40 nm filter with a 535 nm long pass may be more optimal. Due to the excitation and emission characteristics of the acceptor dye, there may be spillover into the PE and PE-CF594 detectors. However, the spillover can be corrected through compensation as with any other dye combination.
Development References (6)
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Godau J, Heller T, Hawlisch H, et al. C5a initiates the inflammatory cascade in immune complex peritonitis.. J Immunol. 2004; 173(5):3437-45. (Clone-specific). View Reference
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Laudes IJ, Chu JC, Huber-Lang M, et al. Expression and function of C5a receptor in mouse microvascular endothelial cells. J Immunol. 2002; 169(10):5962-5970. (Biology). View Reference
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Lee H, Whitfeld PL, Mackay CR. Receptors for complement C5a. The importance of C5aR and the enigmatic role of C5L2. Immunol Cell Biol. 2008; 86(2):153-160. (Biology). View Reference
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Nataf S, Levison SW, Barnum SR. Expression of the anaphylatoxin C5a receptor in the oligodendrocyte lineage. Brain Res. 2001; 894(2):321-326. (Biology). View Reference
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Riedemann NC, Guo RF, Neff TA, et al. Increased C5a receptor expression in sepsis. J Clin Invest. 2002; 110(1):101-108. (Biology). View Reference
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Soruri A, Kim S, Kiafard Z, Zwirner J. Characterization of C5aR expression on murine myeloid and lymphoid cells by the use of a novel monoclonal antibody. Immunol Lett. 2003; 88(1):47-52. (Immunogen). 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.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.