ID | Components | Size |
EP142-C01 | High-bind Plate | 1 plate |
EP142-C02 | Human IL-4 | 25 μg |
EP142-C03 | Biotinylated Human IL-4 Rα | 10 μg |
EP142-C04 | Anti-IL-4Rα Neutralizing Antibody | 20 μg |
EP142-C05 | Streptavidin-HRP | 10 μg |
EP142-C06 | Coating Buffer | 12 mL |
EP142-C07 | 10xWashing Buffer | 50 mL |
EP142-C08 | Blocking Buffer | 50 mL |
EP142-C09 | Substrate Solution | 12 mL |
EP142-C10 | Stop Solution | 7 mL |
背景(Background)
Interleukin-4, is a cytokine that induces differentiation of naive helper T cells (Th0 cells to Th2 cells). In the presence of IL-4 and IL-13, cytokines that are produced in a Th-2 type response, particularly during allergy and parasitic infections, macrophages become differentially activated, And this cytokine is a ligand for interleukin 4 receptor. The interleukin 4 receptor also binds to IL13, which may contribute to many overlapping functions of this cytokine and IL13. STAT6, a signal transducer and activator of transcription, has been shown to play a central role in mediating the immune regulatory signal of this cytokine.
应用说明(Application)
This kit is developed for screening for inhibitors of human IL-4Rα binding to human IL-4.
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 IL-4 Rα pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human IL-4 Rα to immobilized human IL-4 in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinylated IL-4 Rα protein, a human IL-4 protein, an anti-IL-4Rα neutralizing antibody (as method verified Std.), and Streptavidin-HRP reagent.
Your experiment will include 4 simple steps:
a) Coat the plate with human IL-4.
b) Add your molecule of interest to the tests.
c) Add human IL-4 Rα-Biotin to bind the coated IL-4.
d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.
Finally, the half maximal inhibitory concentration (IC50) of your compound to IL-4 Rα: IL-4 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 IL-4Rα [Biotinylated]: IL-4 BINDING BY ANTI-IL-4 Rα NEUTRALIZING ANTIBODY
Serial dilutions of Anti-IL-4 Rα Neutralizing antibody (Catalog # EP142-C04) (1:1 serial dilution, from 5 μg/mL to 0.00244μg/mL) was added into Biotinylated IL-4Rα:IL-4 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).