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 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
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- An isotype control should be used at the same concentration as the antibody of interest.
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.
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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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.
