膜杰作
Star Staining
组分（Materials Provided）
| ID | Components | Size |
| EP168-C01 | High-bind Plate | 1 plate |
| EP168-C02 | Human DR3 | 100 μg |
| EP168-C03 | Biotinylated Human TL1A | 6 μg |
| EP168-C04 | Human Anti-TL1A Neutralizing Antibody | 100 μg |
| EP168-C05 | Streptavidin-HRP | 5 μg |
| EP168-C06 | Coating Buffer | 12mL |
| EP168-C07 | 20xWashing Buffer | 50mL |
| EP168-C08 | Blocking Buffer | 50mL |
| EP168-C09 | Substrate Solution | 12mL |
| EP168-C10 | Stop Solution | 7mL |
背景（Background）
TNF-like cytokine 1A (TL1A) and its receptors, death receptor 3 (DR3) and decoy receptor 3 (DcR3) are members of the TNF and TNF receptor superfamilies of proteins, respectively. Binding of APC-derived TL1A to lymphocytic DR3 provides co-stimulatory signals for activated lymphocytes. DR3 signaling affects not only the proliferative activity of and cytokine production by effector lymphocytes, but also critically influences the development and suppressive function of regulatory T-cells. Whereas, DcR3 restricts the function of the TL1A/DR3 complex: attenuating T-cell activation and downregulating the secretion of pro-inflammatory cytokines. Together with DR3 and DcR3, TL1A constitutes a cytokine system that actively interferes with the regulation of immune responses.
应用说明（Application）
This kit is developed for screening for inhibitors of human TL1A binding to human DR3.
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 Human DR3 pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated Human TL1A to immobilized Human DR3 in a functional ELISA assay and employs a simple colorimetric ELISA platform. Briefly, we provide you with a Human DR3 protein, a Biotinylated Human TL1A, an anti- TL1A neutralizing antibody (as method verified Std.), and Streptavidin-HRP reagent.
Your experiment will include 4 simple steps:
1) Coat the plate with Human DR3.
2) Add your molecule of interest to the tests.
3) Add Human TL1A-Biotin to bind the coated Human DR3.
4) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.
Finally, the half maximal inhibitory concentration (IC50) of your compound to Human TL1A: Human DR3 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.
Serial dilutions of Human Anti-TL1A Neutralizing antibody (Catalog # EP168-C04) (1:1 serial dilution, from 20μg/mL to 0.3125μg/mL) was added into Human DR3: Biotinylated Human TL1A 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).
For each experiment, a standard curve needs to be set for each micro-plate, and the specific OD value may vary depending on different laboratories, testers, or equipments. The example data is for reference only.
背景（Background）：TL1A-DR3
The TL1A protein, belongs to the tumor necrosis factor (TNF) family and is a receptor for TNFRSF25 and TNFRSF6B. TL1A is involved in the activation of NF-κB and C-Jun pathways, which can be used as a regulator of mucosal immunity and participate in the immune pathway of inflammatory bowel disease (IBD) pathogenesis.
