膜杰作
Star Staining
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
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
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
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
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
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
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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.
Permits & Restrictions
This cell line is provided for research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. You are not allowed to share, distribute, sell, modify, sublicense, or otherwise make this cell line available for use to other laboratories, departments, research institutions, hospitals, universities, or biotech companies. 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.
