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. 563794/566349).
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 605 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
- 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.
- CF™ is a trademark of Biotium, Inc.
- Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from BD Horizon™ BV421 may be observed. Therefore, we recommend that individual compensation controls be performed for every BD Horizon™ BV605 conjugate.
Companion Products
The ITEM-4 monoclonal antibody specifically binds to the TWEAK Receptor (TWEAK-R/TWEAKR), which is also known as CD266, or Fibroblast growth factor inducible 14 (FGF-inducible 14, FN14). The TWEAK Receptor is a ~14 kDa type I transmembrane protein that is the 12A family member of the tumor necrosis factor receptor superfamily (TNFRSF12A). It is expressed at low levels in a variety of tissues including, heart, placenta, lung, muscle, and pancreas. TWEAK Receptors are relatively highly expressed on HUVEC cells and certain tumor cell lines. TWEAK-induced TWEAK Receptor-mediated signaling can play roles in inflammation, the induction of apoptosis in certain cell types, the proliferation and migration of endothelial cells, and can promote angiogenesis within healthy and diseased tissues, eg, tumors. The ITEM-4 antibody can induce the death of certain TWEAK-sensitive tumor target cell lines. The TWEAK Receptor binds TWEAK (CD255) and can signal intracellularly by interactions with TRAF1, 2, and 3 cytoplasmic proteins leading to NF-κB activation. The ITEM-4 antibody reportedly crossreacts strongly with mouse CD266 as demonstrated by flow cytometric analysis and functional assays.
This antibody is conjugated to BD Horizon™ BV605 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max of 602-nm, BD Horizon BV605 can be excited by a violet laser and detected with a standard 610/20-nm filter set. BD Horizon BV605 is a tandem fluorochrome of BD Horizon BV421 and an acceptor dye with an Em max at 605-nm. Due to the excitation of the acceptor dye by the green (532 nm) and yellow-green (561 nm) lasers, there will be significant spillover into the PE and BD Horizon PE-CF594 detectors off the green or yellow-green lasers. BD Horizon BV605 conjugates are very bright, often exhibiting brightness equivalent to PE conjugates and can be used as a third color off of the violet laser.
Development References (8)
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Brown SA, Richards CM, Hanscom HN, Feng SL, Winkles JA. The Fn14 cytoplasmic tail binds tumour-necrosis-factor-receptor-associated factors 1, 2, 3 and 5 and mediates nuclear factor-kappaB activation. Biochem J. 2003; 371(Pt 2):395-403. (Biology). View Reference
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Harada N, Nakayama M, Nakano H, Fukuchi Y, Yagita H, Okumura K. Pro-inflammatory effect of TWEAK/Fn14 interaction on human umbilical vein endothelial cells. Biochem Biophys Res Commun. 2002; 299(3):488-493. (Biology). View Reference
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Moreno JA1, Muñoz-García B, Martín-Ventura JL, et al. The CD163-expressing macrophages recognize and internalize TWEAK: potential consequences in atherosclerosis. Atherosclerosis. 2009; 207(1):103-110. (Clone-specific: Western blot). View Reference
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Nakayama M, Harada N, Okumura K, Yagita H. Characterization of murine TWEAK and its receptor (Fn14) by monoclonal antibodies. Biochem Biophys Res Commun. 2003; 306(4):819-825. (Clone-specific: Cytotoxicity, Flow cytometry, Functional assay). View Reference
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Nakayama M, Ishidoh K, Kojima Y, et al. Fibroblast growth factor-inducible 14 mediates multiple pathways of TWEAK-induced cell death. J Immunol. 2003; 170(1):341-348. (Immunogen: Activation, Apoptosis, Cytotoxicity, Flow cytometry, Functional assay, Stimulation). View Reference
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Nakayama M, K. Ishidoh, N. Kayagaki, et al. Multiple pathways of TWEAK-induced cell death. J Immunol. 2002; 168(2):734-743. (Biology). View Reference
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Roos C, Wicovsky A, Müller N, et al. Soluble and transmembrane TNF-like weak inducer of apoptosis differentially activate the classical and noncanonical NF-kappa B pathway. J Immunol. 2010; 185(3):1593-1605. (Clone-specific: Blocking, Functional assay). View Reference
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Wiley SR, Cassiano L, Lofton T, et al. A novel TNF receptor family member binds TWEAK and is implicated in angiogenesis. Immunity. 2001; 15(5):837-846. (Biology). 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.