Mouse PD-L1 / TCR Activator Mammalian Expression Kit

Catalog #

79778

$590 *

Size: 500 reactions

Qty

Bulk/Custom

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*US Pricing only. For international pricing, please contact your local distributor.

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Description

The recombinant expression vectors are designed to express human engineered T cell receptor (TCR) activator and mouse PD-L1 (GenBank Accession #NM_021893) in mammalian cells. The transfected cells can be used in conjunction with mouse PD-1/NFAT Reporter/Jurkat T cells (BPS Bioscience #79762) to study the interactions of PD-1 with PD-L1 ligand in a cellular context and to screen for modulators of this signaling pathway.

Functional Validation and Assay Performance

In this assay, mouse PD-1/NFAT Reporter/Jurkat T cells are used as effector cells; HEK293 cells transiently transfected with the PD-L1 vector over-express mouse PD-L1 and an engineered T cell receptor (TCR) activator and 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 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. Mouse PD1/PD-L1 neutralizing antibodies block PD1:PD-L1 interaction and promote T cell activation, resulting in reactivation of the NFAT responsive luciferase reporter.

Assay Principle

Purchase of this cell line is for research purposes only; commercial use requires a separate license. View the full terms and conditions.

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Synonyms

Programmed Cell Death 1, PDCD1, SLEB2, CD279, mouse pdl1 expression, expression kit, pd-l1, pdl1 mouse

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Product Data Gallery

Product Info

Storage and Usage

Citations

Materials Required But Not Supplied

• HEK293 cell 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.]
• Mouse PD-1/NFAT Reporter Jurkat T cells (BPS Bioscience #79762)
• Opti-MEM I Reduced Serum Medium (life technologies #31985-062)
• Thaw Medium 2: RPMI1640 + 10% FBS + 1% Penicillin/Streptomycin (BPS Bioscience #60184)
• Anti-mouse PD-1 neutralizing antibody: Bioxcell #BP0273, clone#29F.1A12
• Anti-mouse PD-L1 neutralizing antibody: Fisher Scientific #50-146-65, clone#MIH5
• 96-well tissue culture-treated white clear-bottom assay plate
• (optional) NFAT Reporter (Luc) – Jurkat Recombinant Cell Line (BPS Bioscience #60621) as negative control for Mouse PD-1/NFAT Reporter Jurkat T cells (BPS Bioscience #79762)
• ONE-Step™ luciferase assay system (BPS Bioscience #60690) or other luciferase reagents for measuring firefly luciferase activity
• Luminometer

Format

Component

Sepcification

Amount

Storage

TCR activator
+
Mouse PD-L1

(Component A)

Expression vectors constitutively expressing
TCR activator and mouse PD-L1

500 µl
(100 ng/DNA/µl)

-20°C

TCR activator

(Component B)

Expression vector constitutively expressing
TCR activator

500 µl
(100 ng/DNA/µl)

-20°C

Background

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.

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Mouse PD-L1 / TCR Activator Mammalian Expression Kit

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