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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® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
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.
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: Blocking, Flow cytometry, Inhibition, In vivo exacerbation). 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: Blocking, Flow cytometry, Functional assay, Inhibition). 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.