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Application: Useful for screening for inhibitors of TIGIT binding to CD155
Background: Human T-cell immunoreceptor with Ig and ITIM domains (TIGIT) is a receptor that is expressed on the surface of human T cells and NK cells that binds to CD155 and CD112 on the surface of dendritic cells. Binding of TIGIT with CD155 or CD112 results in inhibition of T cell and NK cell activation. Antibodies and other agents that inhibit this signaling pathway have been shown to increase the immune response, especially in the case of certain cancers.
Contraindications: Only limited amounts of DMSO can be included, as it has been shown to disrupt TIGIT:CD155 interaction. Avoid green and blue dyes that absorb light in the AlphaScreen signal emission range (520-620 nm), such as Trypan Blue. Avoid the use of the potent singlet oxygen quenchers such as sodium azide (NaN3) or metal ions (Fe2+, Fe3+, Cu2+, Zn2+ and Ni2+). The presence of >1% RPMI 1640 culture medium leads to a signal reduction due to the presence of excess biotin and iron in this medium. MEM, which lacks these components, does not affect AlphaScreen assays.
Description: The TIGIT:CD155 Homogeneous Assay Kit is designed to measure the inhibition of TIGIT binding to CD155. The TIGIT:CD155 Homogeneous Assay Kit comes in a convenient AlphaLISA® format with purified biotinylated TIGIT, His-tagged CD155, and assay buffer to perform a total of 384 reactions. With this kit, only three simple steps on a microtiter plate are required. First, a sample containing TIGIT and an inhibitor of choice is incubated with the CD155 for 60 minutes. Next, acceptor beads are added, then donor beads, followed by reading the Alpha-counts.
Storage Stability: At least one year from date of receipt when stored as directed.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Yu, X., et al., Nat. Immunol. 2009; 10(1): 48-57.
2. Stanietsky, N., et al., Proc. Natl. Acad. Sci. 2009; 106(42): 17858-17863.
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