TIGIT:CD112 Homogeneous Assay Kit

Catalog #: 72030
Size: 384 reactions
$325
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.
Description:
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.
Synonym(s):
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
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.
Format:

Instructions for use:
See assay kit datasheet for detailed protocol.
Storage / Stability:
At least one year from date of receipt when stored as directed.
Application(s):
Useful for screening for inhibitors of TIGIT binding to CD112
Reference(s):

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.

Notes:

MATERIALS OR INSTRUMENTS REQUIRED BUT NOT SUPPLIED:
AlphaLISA Ni Chelate Acceptor beads, 5 mg/ml (PerkinElmer #AL108C)
AlphaScreen Streptavidin-conjugated Donor beads, 5 mg/ml (PerkinElmer #6760002S)
Optiplate-384 (PerkinElmer #6007290)
AlphaScreen microplate reader
Adjustable micropipettor and sterile tips

Warning(s):
Avoid freeze/thaw cycles.
Scientific Category:
Immunotherapy/Cell Surface Receptor
Product Type:
Assay Kit
Data shown is lot-specific. Contact us for specific information on other lots.

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Product Datasheets

72030.pdf