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Your Position: Home > Kits > TNF-alpha > EP-147

TNF-alpha [Biotinylated] : TNFR2 Inhibitor Screening ELISA Kit

For research use only.

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EP147-C01High-bind Plate1 plate
EP147-C02Human TNFR215 μg
EP147-C03Biotinylated Human TNF-alpha10 μg
EP147-C04Anti-TNF-alpha Neutralizing Antibody20 μg
EP147-C05Streptavidin-HRP10 μg
EP147-C06Coating Buffer12 mL
EP147-C0710xWashing Buffer 50 mL
EP147-C08Blocking Buffer50 mL
EP147-C09Substrate Solution12 mL
EP147-C10Stop Solution7 mL
  • Background
    Tumor necrosis factor alpha (TNFα) is a cytokine produced primarily by monocytes and macrophages. It is found in synovial cells and macrophages in the tissues.The primary role of TNFα is in the regulation of immune cells. TNFα is able to induce apoptotic cell death, to induce inflammation, and to inhibit tumorigenesis and viral replication. Dysregulation of TNFα production has been implicated in a variety of human diseases, including major depression, Alzheimer's disease and cancer. Recombinant TNFα is used as an immunostimulant under the INN tasonermin. TNFα can be produced ectopically in the setting of malignancy and parallels parathyroid hormone both in causing secondary hypercalcemia and in the cancers with which excessive production is associated.
  • Application

    This kit is developed for screening for inhibitors of human TNF-alpha binding to human TNFR2.

    It is for research use only.

  • Reconstitution
    Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
  • Storage
    Unopened kit should be stored at 2°C -8°C upon receiving. Find the expiration date on the outside packaging and do not use reagents past their expiration date.

    The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.

  • Assay Principles
    This inhibitor screening ELISA kit is designed to facilitate the identification and characterization of new TNF-alpha pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human TNF-alpha to immobilized human TNFR2 in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinylated TNF-alpha protein, a human TNFR2 protein, an anti-TNF-alpha neutralizing antibody (as method verified Std.), and Streptavidin-HRP reagent.

    Your experiment will include 4 simple steps:

    a) Coat the plate with human TNFR2.

    b) Add your molecule of interest to the tests.

    c) Add human TNF-alpha-Biotin to bind the coated TNFR2.

    d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.

    Finally, the half maximal inhibitory concentration (IC50) of your compound to TNF-alpha: TNFR2 binding will be determined by comparing OD readings among different experimental groups.

Typical Data Please refer to DS document for The assay protocol.

Serial dilutions of Anti- TNF-alpha Neutralizing antibody (Catalog # EP147-C04) (1:1 serial dilution, from 10 μg/mL to 0.02μg/mL) was added into Biotinylated TNF-alpha: TNFR2 binding reactions. The assay was performed according to the protocol described below. Background was subtracted from data points prior to log transformation and curve fitting (QC tested).

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