LIGHT : HVEM [Biotinylated] Inhibitor Screening ELISA Kit

背景（Background）

Tumor necrosis factor ligand superfamily member 14(LIGHT) is a tumor necrosis factor (TNF) superfamily ligand that regulates T cell immune responses by signaling through the herpes virus entry mediator (HVEM) and the lymphotoxin beta receptor (LTbetaR). LIGHT has emerged as a potent initiator of T cell co-stimulation signals effecting CTL-mediated tumor rejection, allograft rejection and graft versus host disease. Constitutive expression of LIGHT leads to tissue destruction and autoimmune-like disease syndromes.

应用说明（Application）

This kit is developed for screening for inhibitors of human LIGHT binding to human HVEM.

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 LIGHT pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human HVEM to immobilized human LIGHT in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinylated HVEM protein, a human LIGHT protein, an anti-LIGHT neutralizing antibody (as method verified Std.), and Streptavidin-HRP reagent.

Your experiment will include 4 simple steps:

a) Coat the plate with human LIGHT.

b) Add your molecule of interest to the tests.

c) Add human HVEM-Biotin to bind the coated LIGHT.

d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.

Finally, the half maximal inhibitory concentration (IC50) of your compound to LIGHT: HVEM binding will be determined by comparing OD readings among different experimental groups.