ID | Components | Size |
EP140-C01 | High-bind Plate | 1 plate |
EP140-C02 | Human IL-17A | 10 μg |
EP140-C03 | Biotinylated Human IL-17RA | 10 μg |
EP140-C04 | Anti-IL-17A Neutralizing Antibody | 20 μg |
EP140-C05 | Streptavidin-HRP | 10 μg |
EP140-C06 | Coating Buffer | 12 mL |
EP140-C07 | 10xWashing Buffer | 50 mL |
EP140-C08 | Blocking Buffer | 50 mL |
EP140-C09 | Substrate Solution | 12 mL |
EP140-C10 | Stop Solution | 7 mL |
背景(Background)
Interleukin 17 receptor A (IL17RA) is also known as cluster of differentiation w217 (CDw217), is a pro-inflammatory cytokine secreted by activated T-lymphocytes, belong to ubiquitous type I membrane glycoprotein, and binds with low affinity to interleukin 17A (IL17A). IL 17R mRNA exhibits a broad tissue distribution, and has been detected in virtually all cells and tissues tested . IL 17RA associates with IL 17RC to form a signaling receptor complex for IL 17 and IL 17F . Ligand and IL 17RA ligation promotes T cell activation and the production of IL - 6, G-CSF, SCF, and multiple pro-inflammatory chemokines. Defects in IL17RA are the cause of familial candidiasis type 5 (CANDF5).
应用说明(Application)
This kit is developed for screening for inhibitors of human IL-17A binding to human IL-17RA.
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-17A pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human IL-17RA to immobilized human IL-17A in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinylated IL-17RA protein, a human IL-17A protein, an anti-IL-17A neutralizing antibody (as method verified Std.), and Streptavidin-HRP reagent.
Your experiment will include 4 simple steps:
a) Coat the plate with human IL-17A.
b) Add your molecule of interest to the tests.
c) Add human IL-17 RA-Biotin to bind the coated IL-17 A.
d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.
Finally, the half maximal inhibitory concentration (IC50) of your compound to IL-17 RA: IL-17A 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-17A: IL-17RA[Biotinylated] BINDING BY ANTI-IL-17A NEUTRALIZING ANTIBODY
Serial dilutions of Anti-IL-17A Neutralizing antibody (Catalog # EP140-C04) (1:1 serial dilution, from 20 μg/mL to 0.01μg/mL) was added into IL-17A: Biotinylated IL-17RA 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).