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 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 or 566349).
When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values.
For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.
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.
- BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
- Cy is a trademark of GE Healthcare.
Companion Products
The hC3aRZ8 monoclonal antibody specifically binds to the human C3a Receptor (C3aR). C3aR is a seven-transmembrane glycoprotein, G-protein-coupled receptor that is the specific receptor for C3a anaphylatoxin. The C3aR consists of 482 amino acids forming a single polypeptide chain that is encoded by the C3AR1 gene located on chromosome 12 (location 12p13.31). C3aR are expressed by eosinophils, basophils, neutrophils, dendritic cells, monocytes, macrophages, endothelial cells and some T cells. C3a is a bioactive cleavage product released from Complement Component 3 (C3) during complement activation. C3a plays a role in a variety of cellular immune responses as well as being a potent pro-inflammatory agent. In response to bound C3a, this receptor stimulates cellular responses including chemotaxis, granule enzyme release and superoxide anion production and causes increased vascular permeability. In vivo, C3a production can initiate, contribute to, or exacerbate the inflammatory reactions seen in gram-negative bacterial sepsis, trauma, ARDS, ischemic heart disease, post-dialysis syndrome, and several autoimmune diseases including rheumatoid arthritis, lupus erythematosus, and acute glomerulonephritis.
The antibody was conjugated to BD Horizon™ BB700, which is part of the BD Horizon Brilliant™ Blue family of dyes. It is a polymer-based tandem dye developed exclusively by BD Biosciences. With an excitation max of 485 nm and an emission max of 693 nm, BD Horizon BB700 can be excited by the 488 nm laser and detected in a standard PerCP-Cy™5.5 set (eg, 695/40-nm filter). This dye provides a much brighter alternative to PerCP-Cy5.5 with less cross laser excitation off the 405 nm and 355 nm lasers.
Development References (8)
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Ames RS, Li Y, Sarau HM. Molecular cloning and characterization of the human anaphylatoxin C3a receptor. J Biol Chem. 1996; 271(34):20231-20234. (Biology). View Reference
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Crass T, Raffetseder U, Martin U. Expression cloning of the human C3a anaphylatoxin receptor (C3aR) from differentiated U-937 cells. Eur J Immunol. 1996; 26(8):1944-1950. (Biology). View Reference
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Ember JA, Hugli TE. C3a Receptor. In: Oppenheim JJ, Feldmann M, Durum SK. editors in chief, Joost J. Oppenheim, Marc Feldman ; editors, Scott K. Durum .. et al., ed. Cytokine reference : a compendium of cytokines and other mediators of host defense. London: Academic Press; 2001:2173-2181.
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Sacks SH. Complement fragments C3a and C5a: The salt and pepper of the immune response. Eur J Immunol. 2010; 40(3):668-670. (Biology). View Reference
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Soruri A, Kiafard Z, Dettmer C, Riggert J, Köhl J, Zwirner J. IL-4 down-regulates anaphylatoxin receptors in monocytes and dendritic cells and impairs anaphylatoxin-induced migration in vivo.. J Immunol. 2003; 170(6):3306-14. (Immunogen: Flow cytometry, Functional assay). View Reference
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Strainic MG, Liu J, Huang D, et al. Locally produced complement fragments C5a and C3a provide both costimulatory and survival signals to naive CD4+ T cells.. Immunity. 2008; 28(3):425-35. (Biology). View Reference
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Werfel T, Kirchhoff K, Wittmann M, et al. Activated human T lymphocytes express a functional C3a receptor. J Immunol. 2000; 165(11):6599-6605. (Biology). View Reference
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Zwirner J, Gotze O, Begemann G, Kapp A, Kirchhoff K, Werfel T. Evaluation of C3a receptor expression on human leucocytes by the use of novel monoclonal antibodies. Immunology. 1999; 97(1):166-172. (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.