KC-1458

Jurkat-NFAT-Luc2 Cell Line

26431

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Background of Jurkat-NFAT-Luc2 Cell Line

NFAT (nuclear factor of activator T cells) proteins belongs to a family of transcription factors that are very important in T cells activation, differentiation and tolerance. NFAT pathway is activated by Ca2+ and the Ca2+/calmodulin dependent serine phosphatase calcineurin after the engagement of T cell receptor (TCR).

Specifications

Catalog Number	KC-1458
Cell Line Name	Jurkat-NFAT-Luc2 Cell Line
Host Cell Line	Jurkat
Description	Jurkat cell line stable expressing exogenous luciferase gene under the control of NFAT responsive element.
Quantity	One vial of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	70% RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10% FBS+300ug/ml Hygromycin
Selection Marker	Hygromycin B
Morphology	Lymphoblast
Subculture	Split saturated culture 1:4-1:5 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

Jurkat-NFAT-Luc2 cell line was generated using lentivirus method.

Characterization

Figure 1. Activation of Jurkat-NFAT-Luc2 cell line stimulated with CD3/CD28 antibodies or PMA and ionomycin. 20,000 Jurkat-NFAT-Luc2 cell line was seeded into the 96-well plate, and treated with differentstimulus, then readout with Bright-Glo system.

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS, 300μg/ml Hygromycin)in a 37°C water bath.
Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
Incubate the flask at 37°C, 5% CO₂ incubator.
Split saturated culture 1:4-1:5 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI1640 + 20% FBS + 10% DMSO) fresh immediately before use.
Keep the freezing medium on ice and label cryovials.
Transfer cells to a sterile, conical centrifuge tube, and count the cells.
Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
Aliquot 1 mL of the cell suspension into each cryovial.
Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
Transfer vials to liquid nitrogen for long-term storage.

References

1.Macian, F. NFAT proteins: Key regulators of T-cell development and function. Nat. Rev. Immunol. 5, 472–484 (2005).
2.Hogan, P. G., Chen, L., Nardone, J. & Rao, A. Transcriptional regulation by calcium, calcineurin, and NFAT. Genes Dev. 17, 2205–32 (2003).

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.