膜杰作
Star Staining
背景（Background）
Interleukin-23 subunit alpha (IL-23 alpha) can associates with IL12B to form the IL-23 interleukin, a heterodimeric cytokine which functions in innate and adaptive immunity. IL-23 may constitute with IL-17 an acute response to infection in peripheral tissues. IL-23 binds to a heterodimeric receptor complex composed of IL12RB1 and IL23R, activates the Jak-Stat signaling cascade, stimulates memory rather than naive T-cells and promotes production of proinflammatory cytokines. IL-23 induces autoimmune inflammation and thus may be responsible for autoimmune inflammatory diseases and may be important for tumorigenesis.
应用说明（Application）
This kit is developed for screening for inhibitors of human IL-23 binding to human IL-23R.
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 Human IL-23 pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated Human IL-23R to immobilized Human IL-23 in a functional ELISA assay and employs a simple colorimetric ELISA platform. Briefly, we provide you with a Human IL-23A&IL-12B protein, a Biotinylated Human IL-23R, an anti- IL-23A neutralizing antibody (as method verified Std.), and Streptavidin-HRP reagent.
Your experiment will include 4 simple steps:
1) Coat the plate with Human IL-23A&IL-12B.
2) Add your molecule of interest to the tests.
3) Add Human IL-23R-Biotin to bind the coated Human IL-23A&IL-12B.
4) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.
Finally, the half maximal inhibitory concentration (IC50) of your compound to Human IL-23A: Human IL-23R
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.
Serial dilutions of Anti-IL23A Neutralizing antibody (Catalog # EP167-C04) (1:1 serial dilution, from 10μg/mL to 0.000976563μg/mL) was added into Human IL-23A&IL-12B: Biotinylated Human IL-23 R 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).
