ID | Components | Size |
EP139-C01 | High-bind Plate | 1plate |
EP139-C02 | Human IL-17RA | 15μg |
EP139-C03 | Biotinylated Human IL-17A | 10μg |
EP139-C04 | Anti-IL-17A Neutralizing Antibody | 20μg |
EP139-C05 | Streptavidin-HRP | 10μg |
EP139-C06 | Coating Buffer | 12mL |
EP139-C07 | 10xWashing Buffer | 50mL |
EP139-C08 | Blocking Buffer | 50mL |
EP139-C09 | Substrate Solution | 12mL |
EP139-C10 | Stop Solution | 7mL |
背景(Background)
Interleukin-17A (IL17A) is also known as cytotoxic T-lymphocyte-associated antigen 8 (CTLA8), which is a proinflammatory cytokine produced by activated T cells. IL17A can regulate the activities of NF-kappaB and mitogen-activated protein kinases. Also, IL17A can stimulate the expression of IL6 and cyclooxygenase-2 (PTGS2/COX-2), as well as enhance the production of nitric oxide (NO). Furthermore, IL17A has been found both in glycosylated and nonglycosylated forms. High levels of IL-17 are associated with several chronic inflammatory diseases including rheumatoid arthritis, psoriasis and multiple sclerosis.
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-17A to immobilized human IL-17 RA in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinylated IL-17A protein, a human IL-17 RA 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-17 RA.
b) Add your molecule of interest to the tests.
c) Add human IL-17A-Biotin to bind the coated IL-17 RA.
d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.
Finally, the half maximal inhibitory concentration (IC50) of your compound to IL-17A: IL-17 RA 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[Biotinylated]: IL-17RA BINDING BY ANTI-IL-17A NEUTRALIZING ANTIBODY
Serial dilutions of Anti-IL-17A Neutralizing antibody (Catalog # EP139-C04) (1:1 serial dilution, from 10 μg/mL to 0.01μg/mL) was added into Biotinylated IL-17A: 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).