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.
- 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.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
Companion Products
The OX-86 monoclonal antibody specifically binds to the OX-40 antigen (CD134), also known as OX-40L receptor. CD134 is a 50-kDa type-I membrane glycoprotein that belongs to the NGFR/TNFR superfamily. Mouse CD134 is expressed on activated CD4+ and CD8+ T lymphocytes and has been shown to be the sole receptor for the OX-40 Ligand (OX-40L). In the brains of mice with actively induced experimental allergic encephalomyelitis, the expression of CD134 on CD4+ T lymphocytes correlates with disease progression. The OX-40/OX-40L system supplies a costimulatory signal for T-cell proliferation and B-cell proliferation and differentiation. In addition, OX-40 antigen provides a costimulatory signal that induces T cells to proliferate in a CD28-independent manner. In the intact animal, CD134 does not appear to be essential for many T-cell responses, but it seems to play a major role in the pathogenesis of some autoimmune diseases. The OX-86 mAb stains both CD4+ and CD8+ activated T cells, and this expression pattern has been confirmed using OX-40L-Ig fusion protein. CD134 was also detected, using OX-86 mAb, on B cells after stimulation with anti-IgM plus anti-CD40 mAb HM40-3 (Cat. no. 553721). OX-86 mAb does not block binding of OX-40L to OX-40, and it stimulates T-cell proliferation mildly.
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 (8)
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Akiba H, Oshima H, Takeda K, et al. CD28-independent costimulation of T cells by OX40 ligand and CD70 on activated B cells. J Immunol. 1999; 162(12):7058-7066. (Biology). View Reference
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Baum PR, Gayle RB 3rd, Ramsdell F, et al. Molecular characterization of murine and human OX40/OX40 ligand systems: identification of a human OX40 ligand as the HTLV-1-regulated protein gp34. EMBO J. 1994; 13(17):3992-4001. (Biology). View Reference
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Higgins LM, McDonald SA, Whittle N, Crockett N, Shields JG, MacDonald TT. Regulation of T cell activation in vitro and in vivo by targeting the OX40-OX40 ligand interaction: amelioration of ongoing inflammatory bowel disease with an OX40-IgG fusion protein, but not with an OX40 ligand-IgG fusion protein. J Immunol. 1999; 162(1):486-493. (Clone-specific: Western blot). View Reference
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Pippig SD, Pena-Rossi C, Long J, et al. Robust B cell immunity but impaired T cell proliferation in the absence of CD134 (OX40). J Immunol. 1999; 163(12):6520-6529. (Biology). View Reference
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Stuber E, Neurath M, Calderhead D, Fell HP, Strober W. Cross-linking of OX40 ligand, a member of the TNF/NGF cytokine family, induces proliferation and differentiation in murine splenic B cells. Immunity. 1995; 2(5):507-521. (Biology). View Reference
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Weinberg AD, Vella AT, Croft M. OX-40: life beyond the effector T cell stage. Semin Immunol. 1998; 10(6):471-480. (Biology). View Reference
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Weinberg AD, Wegmann KW, Funatake C, Whitham RH. Blocking OX-40/OX-40 ligand interaction in vitro and in vivo leads to decreased T cell function and amelioration of experimental allergic encephalomyelitis. J Immunol. 1999; 162(3):1818-1826. (Biology). View Reference
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al-Shamkhani A, Birkeland ML, Puklavec M, Brown MH, James W, Barclay AN. OX40 is differentially expressed on activated rat and mouse T cells and is the sole receptor for the OX40 ligand. Eur J Immunol. 1996; 26(8):1695-1699. (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.