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NFAT (Luc) Jurkat Reporter Cell Development Service

For research use only.

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

NFAT Assay Principles

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

NFAT FACS

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

NFAT 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

NFAT APPLICATION

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

NFAT APPLICATION

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

NFAT PASSAGE

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

如有相关细胞池需求请联系我们

 

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.

 

前沿进展

 
 
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