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. This will 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 BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. 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).
- BD Horizon Brilliant Ultraviolet 805 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.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
Companion Products
The M75 monoclonal antibody specifically recognizes an epitope in the extracellular N-terminal proteoglycan-like region of Carbonic Anhydrase IX (CA IX). This zinc metalloenzyme is a transmembrane protein that is encoded by CA9 (Carbonic anhydrase 9). This enzyme is expressed in epithelial cells and carcinoma cell lines. It catalyzes the reversible hydration of carbon dioxide and plays a role in various biological processes including respiration, bone resorption, calcification, pH regulation, and cellular adhesion. Carbonic Anhydrase IX may be involved in controlling cellular proliferation and associated with cellular transformation. It is often expressed by tumor cells in response to hypoxia.
Development References (6)
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Mondal UK, Doroba K, Shabana AM, et al. PEG Linker Length Strongly Affects Tumor Cell Killing by PEGylated Carbonic Anhydrase Inhibitors in Hypoxic Carcinomas Expressing Carbonic Anhydrase IX. Int J Mol Sci. 2021; 22:3. (Clone-specific: Western blot). View Reference
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Opavský R, Pastoreková S, Zelník V, et al. Human MN/CA9 gene, a novel member of the carbonic anhydrase family: structure and exon to protein domain relationships.. Genomics. 1996; 33(3):480-7. (Biology). View Reference
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Pastorekova S, Gillies RJ. The role of carbonic anhydrase IX in cancer development: links to hypoxia, acidosis, and beyond.. Cancer Metastasis Rev. 2019; 38(1-2):65-77. (Biology). View Reference
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Pastoreková S, Závadová Z, Kostál M, Babusíková O, Závada J. A novel quasi-viral agent, MaTu, is a two-component system.. Virology. 1992; 187(2):620-6. (Immunogen: Flow cytometry, Immunofluorescence, Western blot). View Reference
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Zat'ovicová M, Tarábková K, Svastová E, et al. Monoclonal antibodies generated in carbonic anhydrase IX-deficient mice recognize different domains of tumour-associated hypoxia-induced carbonic anhydrase IX.. J Immunol Methods. 2003; 282(1-2):117-34. (Clone-specific: Immunofluorescence, Immunohistochemistry). View Reference
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Závada J, Závadová Z, Pastorek J, Biesová Z, Jezek J, Velek J. Human tumour-associated cell adhesion protein MN/CA IX: identification of M75 epitope and of the region mediating cell adhesion.. Br J Cancer. 2000; 82(11):1808-13. (Clone-specific: Blocking, ELISA, Flow cytometry, Functional assay, Western blot). View Reference
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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.