TCR Activator / PD-L2 Mammalian Expression Kit

Catalog #: 60620
Size: 500 reactions
Species: Human
The binding of Programmed Cell Death Protein 1 (PD-1), a receptor expressed on activated T cells, to its ligands, PD-L1 and PD-L2, negatively regulates immune responses. The PD-1 ligands are found on most cancers, and PD-1:PD-L1/2 interaction inhibits T cell activity and allows cancer cells to escape immune surveillance. The PD-1:PD-L1/2 pathway is also involved in regulating autoimmune responses, making these proteins promising therapeutic targets for a number of cancers, as well as multiple sclerosis, arthritis, lupus, and type I diabetes.

The recombinant expression vectors are designed to express human engineered T cell receptor (TCR) activator and human PD-L2 (GenBank Accession #NM_025239) in mammalian cells. The transfected cells can be used in conjunction with PD-1/NFAT Reporter/Jurkat T cells (BPS #60535) to study the interactions of PD-1 with PD-L2 ligand in a cellular context and screen for modulators of this signaling pathway.

In this assay, PD-1/NFAT Reporter/Jurkat T cells are used as effector cells; HEK293 cells overexpressing PD-L1 (or PD-L2) and an engineered T cell receptor (TCR) activator are used as target cells. When these two cells are co-cultivated, TCR complexes on effector cells are activated by TCR activator on target cells, resulting in expression of the NFAT luciferase reporter. However, PD1 and PD-L1 or PD-L2 ligation prevents TCR activation and suppresses the NFAT-responsive luciferase activity. This inhibition can be specifically reversed by anti-PD1 or anti-PD-L1 antibodies. PD1, PD-L1, or PD-L2 neutralizing antibodies block PD1:PD-L1 and/or PD1:PD-L2 interaction and promote T cell activation, resulting in reactivation of the NFAT responsive luciferase reporter.

Programmed Cell Death 2 Ligand 2, PDL2, PD-L2, CD273, PDCDL1G2, B7-DC


Instructions for use:
See assay kit protocol for detailed instructions.
Storage / Stability:
Stable at least 6 months from date of receipt, when stored as directed.
• Screen for activators or inhibitors of PD-1 signaling in a cellular context • Characterize the biological activity of PD-1 and its interactions with ligands
Materials Required but Not Supplied
• HEK293 cells and its growth medium or other cell lines
• Transfection reagent for mammalian cell line [We use Lipofectamine™ 2000 (life technologies #11668027). However, other transfection reagents work equally well.]
• PD-1/NFAT Reporter/Jurkat T cells (BPS Bioscience #60535)
• Opti-MEM I Reduced Serum Medium (life technologies #31985-062)
• Assay medium: RPMI1640 + 10% FBS + 1% Penicillin/Streptomycin
• Anti-PD-1 neutralizing antibody (BPS Bioscience #71120)
• 96-well tissue culture-treated white clear-bottom assay plate
• One-Step luciferase assay system: (BPS Bioscience #60690) or other luciferase reagents for measuring firefly luciferase activity
• Luminometer
Avoid freeze/thaw cycles.
Scientific Category:
Product Type:
Assay Kit
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