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Application: Screen or titrate small molecule inhibitors or biologics for drug discovery and high-throughput screening (HTS) applications of the TL1A binding to DR3.
Background: DR3 (death receptor 3), also known as tumor necrosis factor receptor superfamily member 25 or TNFRSF25, is a membrane receptor of the tumor necrosis factor receptor superfamily of proteins (TNFRSF), which associates with TL1A (TNF-like protein 1A) in T and NK cells. DR3 has been recognized as a significant anti-apoptotic and differentiation factor and it is a co-stimulatory receptor. TL1A, also called TNFSF15, is a member of the tumor necrosis factor family. It is expressed in different immune cells, such as monocyte, macrophage, dendritic cell, T cell and non-immune cells. TL1A competitively binds to DR3, having a higher affinity for DcR3 (decoy receptor 3) providing stimulatory signal for downstream signaling pathways, and then regulates proliferation, activation, apoptosis, and chemokine production in effector cells. The role of DR3 in T cell activation and consequently in cytokine secretion and cell proliferation, make it an attractive target in cancer therapy. Inhibition of DR3-TL1A interaction has substantial therapeutic potential in the treatment of solid tumors.
Contraindications: This kit is compatible with up to 1% final DMSO concentration. Compounds that are fluorescent may interfere with the results, depending on their spectral excitation and emission properties.It is recommended that the compound alone is tested to determine any potential interference of the compound on the assay results.
Description: The TL1A: DR3 Inhibitor Screening TR-FRET Assay Kit is designed for the screening and profiling of neutralizing antibodies or inhibitors of the interaction between TL1A (TNF-like ligand 1A) and DR3 (Death Receptor 3) using TR-FRET (Time-Resolved Fluorescence Resonance Energy Transfer). This kit requires no time-consuming washing steps and comes in a convenient 384-well format, with purified biotinylated-DR3 (amino acids 25-199), europium-labeled (Eu) TL1A (amino acids 72-end), Dye-labeled Acceptor, and assay buffer for 400 reactions. Figure 1. Illustration of the mechanism of the TL1A: DR3 Inhibitor Screening TR-FRET Assay Kit.A sample containing europium-labeled TL1A, a dye-labeled streptavidin acceptor, biotinylated DR3, and an inhibitor, is incubated for 30 minutes. The europium-labeled TL1A binds to DR3-biotin, which receives the dye-labeled streptavidin acceptor. The TR-FRET signal is generated by proximity induced upon interaction of TL1A with DR3. Then, the fluorescence intensity is measured using a fluorescence reader capable of TR-FRET measurements. The signal generated is proportional to TL1A and DR3 binding activity.
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Supplied As: This kit comes in a convenient 384-well format, with purified biotinylated-DR3 (amino acids 25-199), europium-labeled (Eu) TL1A (amino acids 72-end), Dye-labeled Acceptor, and assay buffer for 400 reactions.
Target: DR3, TL1A
Uniprot: O95150, Q93038
Unit Definition: Assay Kit
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: Xu, W. D., et al., 2022 Front. Immunol. 13: 1-10.Zwolak, A., et al., 2022 Sci. Rep. 12(1): 20538.
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