ID | Components | Size |
EP165-C01 | Human TNF-alpha coated Plate | 1 plate |
EP165-C02 | Anti-TNF-alpha Neutralizing Antibody | 10 μg |
EP165-C03 | HRP-Human TNFR1 | 10 μg |
EP165-C04 | 20xWashing Buffer | 50 mL |
EP165-C05 | 1×Dilution Buffer | 50 mL |
EP165-C06 | Substrate Solution | 12 mL |
EP165-C07 | Stop Solution | 7 mL |
背景(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.
应用说明(Application)
This kit is developed for screening for inhibitors of human TNF-alpha binding to human TNFR1.
It is for research use only.
重构方法(Reconstitution)
Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
存储(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.
原理(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 HRP human TNFR1 to immobilized human TNF-alpha in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human HRP TNFR1 protein, a human TNF-alpha 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 TNF-alpha.
b) Add your molecule of interest to the tests.
c) Add HRP-human TNFR1 to bind the coated TNF-alpha.
d) Add TMB or other colorimetric HRP substrate.
Finally, the half maximal inhibitory concentration (IC50) of your compound to TNF-alpha: Osteoprotegerin binding will be determined by comparing OD readings among different experimental groups.
典型数据-Typical Data Please refer to Ds document for the assay protocol.
INHIBITION OF TNF-alpha: TNFR1[HRP] BINDING BY ANTI- TNF-alpha NEUTRALIZING ANTIBODY
Serial dilutions of ANTI- TNF-alpha Neutralizing Antibody (Catalog # EP165-C02) (1:1 serial dilution, from 5μg/mL to 0.002441406 μg/mL) was added into TNF-alpha: TNFR1[HRP] 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).