KC-1548

Jurkat-NFAT-Luc2-LAIR1-CD3-zeta Cell Line

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Background of Jurkat-NFAT-Luc2-LAIR1-CD3-zeta Cell Line

LAIR1 (Leukocyte Associated Immunoglobulin Like Receptor 1) is a Protein Coding gene. Diseases associated with LAIR1 include Malaria and Leukemia, Acute Myeloid. Among its related pathways are Innate Immune System and Class I MHC mediated antigen processing and presentation.

Specifications

Catalog Number	KC-1548
Cell Line Name	Jurkat-NFAT-Luc2-LAIR1-CD3-zeta Cell Line
Host Cell Line	Jurkat-NFAT-Luc2
Description	Stable Jurkat cell line expressing exogenous luciferase under the control of LAIR1-CD3-zeta signaling pathway.
Quantity	Two vials 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% DMEM + 20% FBS + 10% DMSO
Propagation Medium	DMEM + 10% FBS + 300μg/mL Hygromycin B + 0.75μg/mL Puromycin
Selection Marker	Hygromycin B, Puromycin
Morphology	Lymphoblast
Subculture	Split saturated culture 1 : 10 every 3 days; seed out at about 1 × 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-LAIR1-CD3-zeta Cell Line was generated using a lentiviral vector expressing the LAIR1-CD3-zeta sequence.

Characterization

Figure1: Ab1/Ab2/Ab3 were coated overnight and washed twice with PBS, and Jurkat-NFAT-LAIR1-CD3-Zeta cells were seeded in a 96-well plate for 6 hours, and then read out using Bright-Glo Detection System.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS, 300μg/mL Hygromycin B and 0.75μg/mL Puromycin.) 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 : 10 every 3 days; seed out at about 1 × 10⁵ cells/mL

Cell Freezing

Prepare the freezing medium (70% DMEM + 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

Saito F, Hirayasu K, Satoh T, Wang CW, Lusingu J, Arimori T, Shida K, Palacpac NMQ, Itagaki S, Iwanaga S, Takashima E, Tsuboi T, Kohyama M, Suenaga T, Colonna M, Takagi J, Lavstsen T, Horii T, Arase H. Immune evasion of Plasmodium falciparum by RIFIN via inhibitory receptors. Nature. 2017 Dec 7;552(7683):101-105. doi: 10.1038/nature24994. Epub 2017 Nov 29. Erratum in: Nature. 2018 Feb 22;554(7693):554. doi: 10.1038/nature25498. PMID: 29186116; PMCID: PMC5748893.
León-Letelier RA, Dou R, Vykoukal J, Sater AHA, Ostrin E, Hanash S, Fahrmann JF. The kynurenine pathway presents multi-faceted metabolic vulnerabilities in cancer. Front Oncol. 2023 Oct 9;13:1256769. doi: 10.3389/fonc.2023.1256769. PMID: 37876966; PMCID: PMC10591110.
Xu W, Li S, Puan KJ, Li X, Xu C, Fan J, Dou Z, Zhang J, Ju D. Development of an anti-LAIR1 antibody-drug conjugate for acute myeloid leukemia therapy. Int J Biol Macromol. 2025 Mar;293:139432. doi: 10.1016/j.ijbiomac.2024.139432. Epub 2025 Jan 1. PMID: 39753170.