膜杰作
Star Staining
| ID | Components | Size |
| EP128-C01 | High-bind Plate | 1 plate |
| EP128-C02 | Human IL-5 R alpha | 30 μg |
| EP128-C03 | Human IL-5 | 25 μg |
| EP128-C04 | Human IL-5-Biotin | 10 μg |
| EP128-C05 | Streptavidin-HRP | 10 μg |
| EP128-C06 | Coating Buffer | 12 mL |
| EP128-C07 | 10xWashing Buffer | 50 mL |
| EP128-C08 | Blocking Buffer | 50 mL |
| EP128-C09 | Substrate Solution | 12 mL |
| EP128-C10 | Stop Solution | 7 mL |
Background/背景
Interleukin (IL)-5 is produced and secreted by T cells and mast cellsa critical,and is the important regulator of eosinophils and a therapeutic target for asthma. The pharmaceutical inhibition of IL-5 has been shown to reduce eosinophil counts and ameliorate asthmatic symptoms in studies on animal models of allergy as well as in human clinical trials. There are currently three commercially available IL-5 inhibitors in the United States. These include reslizumab (trade name Cinqair), benralizumab (trade name Fasenra), and mepolizumab (trade name Nucala). All three medications have been approved by the FDA for the treatment of severe eosinophilic asthma.
Application/应用
This kit is developed for screening for inhibitors of human IL-5Rα binding to human IL-5.
It is for research use only.
Reconstitution/重构方法
Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
Storage/存储
The unopened kit is stable for 12 months from the date of manufacture if stored at 2°C to 8°C. The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.
Assay Principles/原理
This inhibitor screening ELISA kit is designed to facilitate the identification and characterization of new IL-5 pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human IL-5 to immobilized human IL-5 R alpha in a functional ELISA assay, and employs a simple colorimetric sandwich ELISA platform.
Your experiment will include 4 simple steps:
a) Coat the plate with human IL-5 R alpha.
b) Add your molecule of interest to the tests.
c) Add human IL-5-Biotin to bind the coated human IL-5 R alpha.
d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.
Finally, the ability of your compound to inhibit IL-5: IL-5 R alpha Rbinding will be determined by comparing OD readings among different experimental groups.
Typical Data/典型数据
INHIBITION OF IL-5 [BIOTINYLATED]: IL-5 R ALPHA BINDING BY Human IL-5
Serial dilutions of Human IL-5(Catalog # EP128-C03) (1:1 serial dilution, from 10 μg/mL to 0.020 μg/mL (666.67-0.65 nM)) was added into IL-5 R alpha: IL-5-Biotin binding reactions. The assay was performed according to the above-described protocol. Background was subtracted from data points prior to log transformation and curve fitting (QC tested).
Protocol
