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PE Mouse anti-IRF-7
PE Mouse anti-IRF-7

Analysis of IRF-7 in transfected human epithelial cells.  The 293 fetal kidney cell line was either co-transfected with TBK1 and IRF-7 expression vectors (dashed histogram) or un-transfected (solid histogram, left figure).  After 24 hours, the cells were fixed (BD Cytofix™ buffer, Cat. No. 554655) for 10 minutes at 37°C, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with PE Mouse anti-IRF-7 (Cat. No. 558706).  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.  The specificity of mAb K40-321 was confirmed by western blot using unconjugated antibody on lysates from untransfected (left panel) and TBK1/IRF-7 co-transfected (right panel) 293 cells (right figure).  The transfected IRF-7 protein is identified as a band of 66 kDa in the lysate of co-transfected cells.

Analysis of IRF-7 in transfected human epithelial cells.  The 293 fetal kidney cell line was either co-transfected with TBK1 and IRF-7 expression vectors (dashed histogram) or un-transfected (solid histogram, left figure).  After 24 hours, the cells were fixed (BD Cytofix™ buffer, Cat. No. 554655) for 10 minutes at 37°C, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with PE Mouse anti-IRF-7 (Cat. No. 558706).  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.  The specificity of mAb K40-321 was confirmed by western blot using unconjugated antibody on lysates from untransfected (left panel) and TBK1/IRF-7 co-transfected (right panel) 293 cells (right figure).  The transfected IRF-7 protein is identified as a band of 66 kDa in the lysate of co-transfected cells.

商品详情
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BD Phosflow™
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human IRF-7 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_1645519
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


准备和存储

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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

商品通知

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
558706 Rev. 2
抗体详情
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K40-321

Interferon regulatory factor 7 (IRF-7) is a transcription factor that regulates anti-viral defenses by controlling the induction of type-I interferon (IFN) responses.  IRF-7 expression is induced in lymphoid cells by virus infection, as well as by IFN, lipopolysaccharide, and TNF-α.  IRF-7 responses are initiated by Toll-like receptors (TLR) or the cytoplasmic protein retinoic acid inducible gene I (RIG-I).  Upon TLR activation, it forms cytoplasmic complexes with MyD88, an adaptor in the TLR signaling pathways.  The TLR-dependent and RIG-I-dependent pathways activate kinases, such as IKK-ε and TBK1, that phosphorylate IRF-7 and induce movement of IRF-7-containing complexes to the nucleus, where it preferentially activates IFN-α promoters.

The K40-321 monoclonal antibody recognizes human IRF-7, regardless of phosphorylation status.  Our in-house testing is performed on a cell line that has been co-transfected with TBK1 and IRF-7.  Phosphorylation of IRF-7 in the transfectants requires virus infection or over-expression of a signaling molecule of the RIG-I pathway, such as TBK1.  Phosphorylation of endogenous IRF-7 in untransfected cells has not yet been detected.  We confirmed that mAb K40-321 does not cross-react with TBK1 by Western blot analysis using the purified antibody.  

558706 Rev. 2
格式详情
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
558706 Rev.2
报价单和参考
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研发参考 (8)

  1. Honda K, Yanai H, Mizutani T, et al. Role of a transductional–tanscriptional processor complex involving MyD88 and IRF-7 in Toll-like receptor signaling. Proc Natl Acad Sci U S A. 2004; 101(43):15416-15421. (Biology). 查看参考
  2. Honda K, Yanai H, Negishi H, et al. IRF-7 is the master regulator of type-I interferon-dependent immune responses. Nature. 2005; 434:772-777. (Biology). 查看参考
  3. Hoshino K, Sugiyama T, Matsumoto M, et al. IκB kinase-α is critical for interferon-α production induced by Toll-like receptors 7 and 9. Nature. 2006; 440(7086):949-953. (Biology). 查看参考
  4. Kawai T, Sato S, Ishii KJ, et al. Interferon-α induction through Toll-like receptors involves a direct interaction of IRF7 with MyD88 and TRAF6. Nat Immunol. 2004; 5(10):1061-1068. (Biology). 查看参考
  5. Lin R, Mamane Y, Hiscott J. Multiple regulatory domains control IRF-7 activity in response to virus infection. J Biol Chem. 2000; 275(44):34320-34327. (Biology). 查看参考
  6. Matikainen S, Sirén J, Tissari J, et al. Tumor necrosis factor alpha enhances influenza A virus-induced expression of antiviral cytokines by activating RIG-I gene expression. J Virol. 2006; 80(7):3515-3522. (Biology). 查看参考
  7. Paz S, Sun Q, Nakhaei P, et al. Induction of IRF-3 and IRF-7 phosphorylation following activation of the RIG-I pathway. Cell Mol Biol (Noisy-le-grand). 2006; 52(1):17-28. (Biology). 查看参考
  8. Sharma S, tenOever BR, Grandvaux N, Zhou G-P, Lin R, Hiscott J. Triggering the interferon antiviral response through an IKK-related pathway. Science. 2003; 300:1148-1151. (Biology). 查看参考
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558706 Rev. 2

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.