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Your Position: Home > Kits > IL-6 > EP-146

IL-6 [Biotinylated] : IL-6 R alpha Inhibitor Screening ELISA Kit

For research use only.

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IDComponentsSize
EP146-C01High-bind Plate1 plate
EP146-C02Human IL-6 R alpha15 μg
EP146-C03Biotinylated Human IL-610 μg
EP146-C04Anti-IL-6 Neutralizing Antibody10 μg
EP146-C05Streptavidin-HRP10 μg
EP146-C06Coating Buffer12 mL
EP146-C0710xWashing Buffer 50 mL
EP146-C08Blocking Buffer50 mL
EP146-C09Substrate Solution12 mL
EP146-C10Stop Solution7 mL
  • Background
    Interleukin 6 (IL-6) is also known as HGF, BSF2, HSF, IFNB2 and IL-6, originally identified as a B cell differentiation factor, is a multifunctional cytokine that regulates immune responses, hematopoiesis, acute phase responses, and inflammatory reactions.It is secreted by T cells, macrophages, monocytes, fibroblasts, endothelial cells, et.al. to stimulate immune response to trauma, especially burns or other tissue damage leading to inflammation. Interleukin 6 has been shown to interact with interleukin-6 receptor and glycoprotein. IL-6 is relevant to many disease processes such as diabetes, atherosclerosis, depression, Alzheimers Disease, systemic, lupus erythematosus, prostate cancer and rheumatoid arthritis. Advanced/metastatic cancer patients have higher levels of IL-6 in their blood.Hence there is an interest in developing anti-IL-6 agents as therapy against many of these diseases.
  • Application

    This kit is used as a tool to aid in optimal purification process development and in routine quality control of in-process streams as well as final product by various biotechnological processes such as cell or tissue culture.

    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 IL-6 pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human IL-6 to immobilized human IL-6 R alpha in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinylated IL-6 protein, a human IL-6 R alpha protein, an anti-IL-6 neutralizing antibody (as method verified Std.), and Streptavidin-HRP reagent.

    Your experiment will include 4 simple steps:

    a) Coat the plate with human IL-6 R alpha.

    b) Add your molecule of interest to the tests.

    c) Add human IL-6-Biotin to bind the coated IL-6 R alpha.

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

    Finally, the half maximal inhibitory concentration (IC50) of your compound to IL-6: IL-6 R alpha binding will be determined by comparing OD readings among different experimental groups.

Typical Data Please refer to DS document for The assay protocol.
 IL-6 TYPICAL DATA

INHIBITION OF IL-6 [Biotinylated]: IL-6 R alpha BINDING BY ANTI-IL6 NEUTRALIZING ANTIBODY
Serial dilutions of Anti-IL-6 Neutralizing antibody (Catalog # EP146-C04) (1:1 serial dilution, from 10 μg/mL to 0.01μg/mL) was added into Biotinylated IL-6: IL-6 R alpha 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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