The TIGIT:CD112 Homogeneous Assay Kit is designed to measure the inhibition of human T-cell immunoreceptor with Ig and ITIM domains (TIGIT) binding to CD112 (PVRL2/Nectin-2). The TIGIT:CD112 Homogeneous Assay Kit comes in a convenient AlphaLISA® format with purified biotinylated TIGIT, His-tagged CD112, 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 CD112 for 60 minutes. Next, acceptor beads are added, then donor beads, followed by reading the Alpha-counts.
T-cell immunoreceptor with Ig and ITIM domains, V-set and immunoglobulin domain-containing protein 9 (VSIG9), and V-set and transmembrane domaincontaining protein 3 (VSTM3), PVRL2
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.
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.
Storage/Stability
At least 6 months from date of receipt when stored as directed.
Instructions for Use
See assay kit datasheet for detailed protocol.
Applications
Useful for screening for inhibitors of TIGIT binding to CD112
Shipping Temperature
dry ice
Notes
AlphaScreen® and AlphaLISA® are registered trademarks of PerkinElmer, Inc.
Contraindications
Only limited amounts of DMSO can be included, as it has been shown to disrupt TIGIT:CD112 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.
Warnings
Avoid freeze/thaw cycles.
Disclaimers and Limitations
BPS Bioscience assay kits are validated using the listed components according to our specific protocol. Any deviations to kit components or protocols, such as using different proteins, cell/tissue lysates, or buffers (including using the same component from other commercial sources) will void any warranties on the performance of the assay kit and is not recommended.
End-users should immediately report any issues to [email protected] upon receipt of the kit, such as missing components, damaged vials, no dry ice, etc.
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