DESCRIPTION

• 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
• Shiptemp: -80°C (dry ice)
• Synonyms: DNAX accessory molecule 1 (DNAM-1), Poliovirus receptor, PVR, and Nectin-like protein 5
• Category: Immunotherapy/Assay Kit
• Background: CD226 is an activating receptor expressed on the surface of natural killer (NK) cells, monocytes, platelets, and T cells. Its natural ligands are CD112 and CD155 which are expressed by antigen presenting cells (APC). Binding of CD226 to either CD112 or CD155 triggers NK cell effector function and can promote Th1 cell differentiation in T cells. CD226 competes for binding with CD112 and CD155 with the co-inhibitory receptor TIGIT. Agonistic CD226 monoclonal antibodies are being explored as potential therapeutics in settings where immune activation is beneficial, such as cancer.
• References: 1. Bottino, C. et al., J. Exp. Med. 2003; 198(4): 557-567. 2. Pende, D. et al., Blood. 2005; 105(5): 2066-2073.
• Description: The CD226:CD155 Homogeneous Assay Kit is designed to measure the inhibition of CD226 binding to CD155 (PVR). The CD226:CD155 Homogeneous Assay Kit comes in a convenient AlphaLISA® format with purified biotinylated CD226, 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 CD226 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.
• Methodology: AlphaLISA®
• Unspsc Code: 41121811
• Unspsc Name: Laboratory microchemistry kits
• Applications: Useful for screening for inhibitors of CD226 binding to CD155
• Product Type: Assay Kit
• Biosafety Level: Not applicable (BSL-1)
• Related Products: 72029, 72030, 72051
• Contraindications: Only limited amounts of DMSO can be included, as it has been shown to disrupt CD226: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.
• Storage Stability: Stable for at least one year from date of receipt when stored as directed.
• Scientific Category: Immunotherapy/Cell Surface Receptor
• Instructions for Use: See product datasheet for detailed protocol.