膜杰作
Star Staining
描述（Description）
The NFAT (Luc) Jurkat Reporter Cell was engineered with the NFAT response element driving luciferase expressing systems. We could equip this reporter cell with a chimeric antigen receptor (CAR) for developing a CAR-J-based activity screening system. The anti-TCR/CD3 inducing intracellular signals could be inhibited by some transfected immune checkpoints binding to corresponding ligands.
应用说明（Application）
1. Transfection host for some immune checkpoint concerning the NFAT signaling pathway
2. The discovery of T cell activators by the NFAT signaling bioactivity
生长特性（Growth Properties）
Suspension
筛选标记（Selection Marker）
Puromycin (5 μg/mL)
培养基（Culture Medium）
RPMI-1640 medium + 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
Expression analysis of human CD3 on NFAT (Luc) Jurkat Reporter Cell by FACS.
NFAT (Luc) Jurkat Reporter Cell were stained with APC-R700 labeled Anti-Human CD3 antibody or APC-R700 labeled Isotype antibody.
Protocol
Application
Activating of NFAT signaling bioactivity by anti-human CD3 antibody (RLU).
This reporter cell was incubated with serial dilutions of anti-human CD3 antibody (AcroBiosystems, Cat.No.CDE-M120a). The EC50 of anti-human CD3 antibody was approximately 0.078 μg/mL.
Protocol
Activating of NFAT signaling bioactivity by anti-human CD3 antibody (Fold).
This reporter cell was incubated with serial dilutions of anti-human CD3 antibody (AcroBiosystems, Cat.No.CDE-M120a). The max induction fold was approximately 137.
Protocol
Bioactivity detection of Anti-human CD3xCD19 bispecific antibody.
This reporter cell was incubated with serial dilutions of Blinatumomab (CD3×CD19 BsAb) in the presence of Raji cells that express human CD19 endogenously. The EC50 of Blinatumomab incubated with Raji cells is approximately 0.66 pM with the max induction fold 842.
Protocol
Passage Stability
Passage stability analysis by anti-human CD3 antibody stimulation.
The continuously growing NFAT (Luc) Jurkat Reporter Cell was stimulated with serial dilutions of anti-human CD3 antibody (AcroBiosystems, Cat.No.CDE-M120a). Anti-human CD3 antibody stimulated response demonstrates passage stabilization (fold induction and EC50) across passage 15-34.
Protocol
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背景（Background）
Nuclear factor of activated T cells (NFAT), which is the pharmacological target of immunosuppressants cyclosporine and tacrolimus, has been shown to play an important role in T cells (immune system) and many biological events. Accumulating studies have indicated that NFATs are involved in many aspects of cancer, including carcinogenesis, cancer cell proliferation, invasion, metastasis, angiogenesis, drug resistance and tumor microenvironment. As the key section of the adaptive immune response, the T cell receptor (TCR)-calcium-calcineurin signaling pathway could lead to T cell activation. The 'nuclear factor of activated T cells' proteins are transcription factors that promote expression of a panel of genes required for activation.
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.
