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 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
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. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
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.
- 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 750 is covered by one or more of the following US patents: 8,158,444; 8,802,450; 8,575,303; 8,455,613; 8,227,187; 8,841,072; 8,110,673.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The OX-38 antibody specifically recognizes the CD4 antigen on most thymocytes, a subpopulation of mature T lymphocytes (ie, MHC class II-restricted T cells, including most T helper cells), monocytes, macrophages, and some dendritic cells. CD4 is an antigen coreceptor on the T-cell surface which interacts with MHC class II molecules on antigen-presenting cells. It participates in T-cell activation through its association with the T-cell receptor complex and protein tyrosine kinase lck. The OX-38 antibody has been reported to bind to the same epitope of CD4 as that recognized by W3/25 mAb, which is a different epitope than that recognized by OX-35 mAb. In vivo blocking of some cell-mediated immune responses by mAb OX-38 has been reported. Injection of OX-38 mAb induces allograft unresponsiveness in rats, with varying results depending on the rat strain used (high or low responder). Furthermore, in vivo depletion of CD4+ lymphocytes has been reported with this antibody.
The antibody was conjugated to BD Horizon BV750 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 750-nm. BD Horizon Brilliant BV750 can be excited by the violet laser (405 nm) and detected with a 750/30 nm filter with a 740 nm long pass. Due to spectral differences between labeled cells and beads, using BD™ CompBeads can result in incorrect spillover values when used with BD Horizon BV750 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended.
Development References (9)
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Arima T, Goss JA, Walp LA, Flye MW. Administration of anti-CD4 monoclonal antibody with intrathymic injection of alloantigen results in rat cardiac allograft tolerance. Surgery. 1995; 118(2):265-273. (Clone-specific: Depletion). View Reference
-
Bañuls MP, Alvarez A, Ferrero I, Zapata A, Ardavin C. Cell-surface marker analysis of rat thymic dendritic cells. Immunology. 1993; 79(2):298-304. (Biology). View Reference
-
Bierer BE, Sleckman BP, Ratnofsky SE, Burakoff SJ. The biologic roles of CD2, CD4, and CD8 in T-cell activation. Annu Rev Immunol. 1989; 7:579-599. (Biology). View Reference
-
Janeway CA Jr. The T cell receptor as a multicomponent signalling machine: CD4/CD8 coreceptors and CD45 in T cell activation. Annu Rev Immunol. 1992; 10:645-674. (Biology). View Reference
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Jefferies WA, Green JR, Williams AF. Authentic T helper CD4 (W3/25) antigen on rat peritoneal macrophages. J Exp Med. 1985; 162(1):117-127. (Immunogen: Flow cytometry, Functional assay, Immunoaffinity chromatography, Immunoprecipitation, Inhibition). View Reference
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Liu L, Zhang M, Jenkins C, MacPherson GG. Dendritic cell heterogeneity in vivo: two functionally different dendritic cell populations in rat intestinal lymph can be distinguished by CD4 expression. J Immunol. 1998; 161(3):1146-1155. (Biology). View Reference
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Stitz L, Sobbe M, Bilzer T. Preventive effects of early anti-CD4 or anti-CD8 treatment on Borna disease in rats. J Virol. 1992; 66(6):3316-3323. (Clone-specific: Blocking). View Reference
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Suzuki H, Hara MH, Miyahara T, et al. Microchimerism and graft acceptance: IV. Cardiac allograft acceptance following anti-adhesion molecule antibody therapy. Transplant Proc. 1996; 28(4):2058-2060. (Clone-specific: Blocking). View Reference
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Yin D, Fathman CG. Tissue-specific effects of anti-CD4 therapy in induction of allograft unresponsiveness in high and low responder . Transpl Immunol. 1995; 3(3):258-264. (Clone-specific: Blocking). 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.