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BUV661 Mouse Anti-Human CD266
Product Details
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BD OptiBuild™
TWEAKR; TWEAK-R; TWEAK Receptor; TNFRSF12A; FGF-inducible 14; FN14
Human (Tested in Development)
Mouse IgG1, κ
Human
Flow cytometry (Qualified)
0.2 mg/ml
VIII 80430
AB_2874853
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon BUV661 under optimal conditions that minimize unconjugated dye and antibody.

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

  1. This antibody was developed for use in flow cytometry.
  2. 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.
  3. Researchers should determine the optimal concentration of this reagent for their individual applications.
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  8. 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.
  9. BD Horizon Brilliant Ultraviolet 661 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
750734 Rev. 4
Antibody Details
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ITEM-1

The ITEM-1 monoclonal antibody specifically binds to CD266, which is also known as TWEAK Receptor (TWEAK-R/TWEAKR), or Fibroblast growth factor inducible 14 (FGF-inducible 14, FN14). CD266 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. CD266 is relatively highly expressed on HUVEC cells and certain tumor cell lines. TWEAK-induced CD266-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-1 antibody can reportedly induce the proliferation of HUVEC cells and the death of certain TWEAK-sensitive tumor target cell lines. CD266 binds TWEAK (CD255) and can signal intracellularly by interactions with TRAF1, 2, and 3 cytoplasmic proteins leading to NF-κB activation.

The antibody was conjugated to BD Horizon™ BUV661 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 661-nm. BD Horizon Brilliant BUV661 can be excited by the ultraviolet laser (355 nm) and detected with a 670/25 filter and a 630 nm LP.  Due to cross laser excitation of this dye, there may be significant spillover into channels detecting APC-like emissions (eg, 670/25-nm filter).

    

Due to spectral differences between labeled cells and beads, using BD™ CompBeads can result in incorrect spillover values when used with BD Horizon BUV661 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different BUV661 reagents (eg, CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone-specific compensation controls when using these reagents.

750734 Rev. 4
Format Details
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BUV661
The BD Horizon Brilliant™ Ultraviolet 661 (BUV661) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This tandem fluorochrome is comprised of a BUV395 donor with an excitation maximum (Ex Max) of 350-nm and an acceptor dye with an emission maximum (Em Max) at 660-nm. BUV661, driven by BD innovation, is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 660-nm (e.g., 670/25 bandpass filter). The acceptor dye can be excited by the Red (628–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
BUV661
Ultraviolet 355 nm
350 nm
660 nm
750734 Rev.4
Citations & References
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Development References (7)

  1. 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
  2. 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
  3. 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. (Biology). View Reference
  4. 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
  5. 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
  6. 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
  7. 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
View All (7) View Less
750734 Rev. 4

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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.