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Human PD-1/LAG-3 (Luc) Jurkat Reporter cell Development Service

For research use only.

描述(Description)

The Human PD-1/LAG-3 (Luc) Jurkat Reporter Cell was engineered to not only express the NFAT response element driving luciferase expressing systems, but also express the receptors full length human PD-1 (Gene ID: 5133) and LAG-3 (Gene ID: 3902), which can use to evaluate the synergistic effect of anti-human PD-1 and anti-human LAG-3 antibody. When co-cultured with target cells expressing human PD-L1 and MHCⅡ, the PD-1/PD-L1 and LAG-3/MHCⅡ interactions inhibit TCR signaling and NFAT-mediated luminescence. Blocking the PD-1/PD-L1 and LAG-3/MHCⅡ interactions by the stimultaneous addition of anti-PD-1 or anti-PD-L1 and anti-LAG-3 antibodies release the inhibitory signals and result in TCR activation and NFAT-mediated luminescence.

应用说明(Application)

Screen for anti-human PD-1 or/and anti-human LAG-3 antibody

PD1/LAG3 Assay Principles

生长特性(Growth Properties)

Suspension

筛选标记(Selection Marker)

Puromycin (5 μg/mL) + Hygromycin (20 μg/mL)

培养基(Culture Medium)

RPMI-1640 + 10% FBS

冻存液(Freeze Medium)

Serum-free cell cryopreservation medium

装量(Quantity)

1 vial contains at least 5×10^6 cells in 1 mL serum-free cryopreservation medium

存储(Storage)

Frozen in liquid nitrogen.

支原体检测(Mycoplasma Testing)

Negative

无菌检测(Sterility Testing)

Negative

使用说明(Instructions for Use)

See data sheet for detailed culturing and assay protocol.

 

Receptor Assay

PD1/LAG3 FACS

Co-expression analysis of human PD-1 and LAG-3 on Human PD-1/LAG-3 (Luc) Jurkat Reporter Cell by FACS.
Cell surface staining was performed on Human PD-1/LAG-3 (Luc) Jurkat Reporter Cell or negative control cell using FITC-labeled anti-human PD-1 antibody and APC-labeled anti-human LAG-3 antibody.

Protocol

 

Application

PD1/LAG3 APPLICATION

Analysis of the synergistic effect for anti-human PD-1 and anti-human LAG-3 antibody (RLU).
This reporter cell was co-incubated with serial dilutions of anti-human PD-1 plus anti-human LAG-3 antibody in the presence of target cells expressing human PD-L1 and MHCⅡ. The EC50 was approximately 0.58 μg/mL.

Protocol

PD1/LAG3 APPLICATION

Analysis of the synergistic effect for anti-human PD-1 and anti-human LAG-3 antibody (FOLD).
This reporter cell was co-incubated with serial dilutions of anti-human PD-1 plus anti-human LAG-3 antibody in the presence of target cells expressing human PD-L1 and MHCⅡ. The max induction fold was approximately 40.

Protocol

 

Passage Stability

PD1/LAG3 PASSAGE

Passage stability analysis by Signaling Bioassay.
The continuously growing Human PD-1/LAG-3 Jurkat Reporter Cell was stimulated with serial dilutions of anti-human PD-1 plus anti-human LAG-3 antibody in the presence of target cells expressing PD-L1 and MHCⅡ. Anti-human PD-1 plus anti-human LAG-3 antibody stimulated response demonstrates passage stabilization (fold induction and EC50) across passage 8-25.

Protocol

 

背景(Background)

Programmed cell death protein 1 (PD-1) is also known as CD279 and PDCD1, is a type I membrane protein and is a member of the extended CD28/CTLA-4 family of T cell regulators. PDCD1 is expressed on the surface of activated T cells, B cells, macrophages, myeloid cells and a subset of thymocytes. PD1 inhibits the T-cell proliferation and production of related cytokines including IL-1, IL-4, IL-10 and IFN-γ by suppressing the activation and transduction of PI3K/AKT pathway.

Lymphocyte activation gene 3 protein (LAG3) is also known as CD antigen CD223 and protein FDC, which belongs to immunoglobulin (Ig) superfamily. As a CD4 homologue, LAG3 expressed on the surface of activated conventional T cells and regulatory T (Treg) cells. In conventional T cells, LAG-3 acts as an inhibitory receptor of T cell inflammation.

 

License Disclosure

This cell line is provided for research use only. This license does not permit you to share, distribute, sell, sublicense, or otherwise make this cell line available for use to other laboratories, departments, research institutions, hospitals, universities, or biotech companies. The license does not permit modification of this cell line in any way. Inappropriate use or distribution of this cell line will result in revocation of the license. Modifications of this cell line, transfer to another facility, or commercial use of the cell lines may require a separate license and additional fees. AcroBiosystems does not warrant the suitability of this cell line for any particular use, and does not accept any liability in connection with the handling or use of this cell line.

 

前沿进展

 
 
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