膜杰作
Star Staining
组分（Materials Provided）
| ID | Components | Size |
| EP155-C01 | High-bind Plate | 1 plate |
| EP155-C02 | Human Integrin α 4 β 7 | 25 μg |
| EP155-C03 | Biotinylated Human MAdCAM-1 | 10 μg |
| EP155-C04 | Anti-Integrin α 4 β 7 Neutralizing Antibody | 20 μg |
| EP155-C05 | Streptavidin-HRP | 5 μg |
| EP155-C06 | Coating Buffer | 12 mL |
| EP155-C07 | 10xWashing Buffer | 50 mL |
| EP155-C08 | Blocking Buffer | 50 mL |
| EP155-C09 | Substrate Solution | 12 mL |
| EP155-C10 | Stop Solution | 7 mL |
| EP155-C11 | 1mol/L CaCl₂ | 1.5 mL |
| EP155-C12 | 1mol/L MgCl₂ | 1.5 mL |
| EP155-C13 | 1mol/L MnCl₂ | 1.5 mL |
背景（Background）
The integrin α4β7 is heterodimeric cell surface receptors expressed on most leukocytes. Mucosal addressing cell adhesion molecule 1(MAdCAM-1) is an exclusive ligand for α4β7 integrin. The interaction of integrin α4β7 with addressin mucosal addressin cell adhesion molecule-1 (MAdCAM-1) is central to IBD pathogenesis.
应用说明（Application）
This kit is developed for screening for inhibitors of human Integrin α 4 β 7 binding to human MAdCAM-1.
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 Integrin α 4 β 7 pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human MAdCAM-1 to immobilized human Integrin α 4 β 7 in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinylated MAdCAM-1 protein, a human Integrin α 4 β 7 protein, an anti-Integrin α 4 β 7 neutralizing antibody (as method verified Std.), and Streptavidin-HRP reagent.
Your experiment will include 4 simple steps:
a) Coat the plate with human Integrin α 4 β 7.
b) Add your molecule of interest to the tests.
c) Add human MAdCAM-1-Biotin to bind the coated Integrin α 4 β 7.
d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.
Finally, the half maximal inhibitory concentration (IC50) of your compound to Integrin α 4 β 7:MAdCAM-1 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 Integrin α 4 β 7: MAdCAM-1 [Biotinylated] BINDING BY ANTI-Integrin α 4 β 7 NEUTRALIZING ANTIBODY
Serial dilutions of Anti-Integrin α 4 β 7 Neutralizing antibody (Catalog # EP155-C04) (1:1 serial dilution, from 5 μg/mL to 0.0098μg/mL) was added into Integrin α 4 β 7: Biotinylated MAdCAM-1 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).
