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Your Position: 홈 > Kits > TNF-alpha > EP-143

TNF-alpha [Biotinylated] : TNFR1 Inhibitor Screening ELISA Kit

For research use only.
IDComponentsSize
EP143-C01High-bind Plate1 plate
EP143-C02Human TNFR115 μg
EP143-C03Biotinylated Human TNF-alpha10 μg
EP143-C04Anti-TNF-alpha Neutralizing Antibody20 μg
EP143-C05Streptavidin-HRP10 μg
EP143-C06Coating Buffer12 mL
EP143-C0710xWashing Buffer 50 mL
EP143-C08Blocking Buffer50 mL
EP143-C09Substrate Solution12 mL
EP143-C10Stop Solution7 mL
  • Background
    Tumor Necrosis Factor (TNF), also known as TNFα, is a kind of cytokine that can kill tumor cells directly and has no obvious cytotoxicity to normal cells. TNF is a cell signaling protein involved in systemic inflammation, and it is one of the cytokines that constitute the acute phase response. It is produced primarily by activated macrophages, but most other immune cells, such as CD4+ lymphocytes, NK cells, neutrophils, mast cells, eosinophils, and neurons in the nervous system can also produce it.
  • Application

    This kit is developed for screening for inhibitors of human TNF-alpha binding to human TNFR1.

    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 TNFR1 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 TNFR1 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 TNFR1 .

    b) Add your molecule of interest to the tests.

    c) Add human TNF-alpha-Biotin to bind the coated TNFR1.

    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: TNFR1 binding will be determined by comparing OD readings among different experimental groups.

Typical Data Please refer to Ds document for the assay protocol.
TNF-alpha TYPICAL DATA

INHIBITION OF TNF-alpha [Biotinylated]: TNFR1 BINDING BY ANTI- TNF-alpha NEUTRALIZING ANTIBODY
Serial dilutions of Anti-TNF-alpha Neutralizing antibody (Catalog # EP143-C04) (1:1 serial dilution, from 10 μg/mL to 0.005μg/mL) was added into Biotinylated TNF-alpha: TNFR1 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).

  • Clinical and Translational Updates

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