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.
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).
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- 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).
- 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 DY12 monoclonal antibody specifically recognizes CD245 which is also known as p220/240. CD245 is variably expressed by T cells, B cells. NK cells, granulocytes, and platelets and more highly expressed on monocytes. CD245 expression can be upregulated on activated T cells, B cells, and NK cells. CD245 has been identified as Myosin 18A that is encoded by MYO18A (Myosin XVIIIA). Myosin 18A is also known as Myosin containing a PDZ domain (MysPDZ), Molecule associated with JAK3 N-terminus (MAJN), and Surfactant protein receptor SP-R2101 (SP-R2101). Myosin 18A is involved in intracellular transport processes and can also reportedly serve as a cell surface receptor for Surfactant Protein A (SP-A). The DY12 antibody reportedly immunoprecipitated long α (230 kDa) and short β (190 kDa) isoforms of Myosin 18A expressed by normal human lung extracts and by the YT2C2 NK cell line-further evidence suggested that the Myosin 18Aα isoform is expressed at the cell surface. CD245 stimulation mediated by bound and crosslinked DY12 antibody or the SP-A ligand can reportedly augment NK cell degranulation or IL-2-activated NK cell-mediated cytotoxicity against certain tumor target cells. Moreover, stimulation through CD245 can upregulate CD137/4-1BB expression on NK cells and promote NK cell-mediated cytotoxicity against CD137 Ligand-expressing target cells.
The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.
Development References (5)
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Boumsell L, Nikolova M, Marie-Cardine A, Bensussan A. CD245 Workshop report. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:691-692.
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De Masson A, Giustiniani J, Marie-Cardine A, et al. Identification of CD245 as myosin 18A, a receptor for surfactant A: A novel pathway for activating human NK lymphocytes.. Oncoimmunology. 2016; 5(5):e1127493. (Immunogen: Flow cytometry, Functional assay, Immunohistochemistry). View Reference
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Gurka S, Dirks S, Photiadis J, Kroczek RA. Expression analysis of surface molecules on human thymic dendritic cells with the 10th HLDA Workshop antibody panel.. Clin Transl Immunology. 2015; 4(10):e47. (Clone-specific: Flow cytometry). View Reference
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Okumura K, Kato K, Kobata T. T-cell antigens: Section report. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:23-32.
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Zola H, Swart B, Boumsell L, Mason DY. Human Leucocyte Differentiation Antigen nomenclature: update on CD nomenclature. Report of IUIS/WHO Subcommittee.. J Immunol Methods. 2003; 275(1-2):1-8. (Clone-specific). View Reference
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