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Applications: This kit is useful for screening for inhibitors of VISTA binding to VSIG-3.
Background: V-domain Ig Suppressor of T cell activation (VISTA, or B7-H5) is a type I transmembrane protein that functions as an immune checkpoint. V-Set and Immunoglobulin domain containing 3 (VSIG-3, or IGSF11) is a ligand for VISTA which, when bound on activated T cells, inhibits their proliferation and the production of cytokines and chemokines. VSIG-3 expression is elevated in many cancers, especially glioma and melanoma, so the blockade of the VSIG-3/VISTA pathway represents a promising potential cancer immunotherapy strategy.
Contraindications: Keep final DMSO concentration at or below 1%. Excess DMSO interferes with binding and decreases sensitivity of the assay.
Description: The VSIG-3:VISTA[Biotinylated] Inhibitor Screening Assay Kit is designed for screening and profiling inhibitors of VISTA binding to VSIG-3. This kit comes in a convenient 96-well format, with biotin-labeled VISTA, purified VSIG-3, streptavidin-labeled HRP, and assay buffer for 100 binding reactions. The key to this kit is the high sensitivity of detection of biotin-labeled VISTA by streptavidin-HRP. Only a few simple steps on a microtiter plate are required for the assay. First, VSIG-3 is coated on a 96-well plate. Next, biotinylated VISTA is incubated with VSIG-3 on the plate. Finally, the plate is treated with streptavidin-HRP followed by addition of an HRP substrate to produce chemiluminescence, which can be measured using a chemiluminescence reader.
Storage Stability: Up to 6 months from date of receipt, when stored as recommended.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: Wang, J., et al. Immunol. 2019, 156.1: 74-85.
Xu, W., et al. Cell. Mol. Immunol. 2018, 15(5): 438-446.
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