KC-5341

293T-NFκB-Luc2-mouse-HVEM Cell Line

55776

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Background of 293T-NFκB-Luc2-mouse-HVEM Cell Line

This gene encodes a member of the TNF (tumor necrosis factor) receptor superfamily. The encoded protein functions in signal transduction pathways that activate inflammatory and inhibitory T-cell immune response. It binds herpes simplex virus (HSV) viral envelope glycoprotein D (gD), mediating its entry into cells. Alternative splicing results in multiple transcript variants.TNFRSF14 (TNF Receptor Superfamily Member 14) is a Protein Coding gene. Diseases associated with TNFRSF14 include Herpes Simplex and Leukemia, Chronic Lymphocytic. Among its related pathways are Akt Signaling and TNF Superfamily - Human Ligand-Receptor Interactions and their Associated Functions. Gene Ontology (GO) annotations related to this gene include ubiquitin protein ligase binding and tumor necrosis factor receptor activity.

Specifications

Catalog Number	KC-5341
Cell Line Name	293T-NFκB-Luc2-mouse-HVEM Cell Line
Clone Number	4A3
Host Cell Line	293T-NFκB-Luc2
Description	Stable Jurkat cell line expressing exogenous luciferase under the control of NFκB responsive element and mouse HVEM fusion sequence.
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% DMEM + 20% FBS + 10% DMSO
Propagation Medium	DMEM + 10% FBS + 150μg/mL Hygromycin B + 1μg/mL puromycin
Selection Marker	Hygromycin B and Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:6 every 2-3 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

Cell Line Generation

293T-NFκB-Luc2-mouse-HVEM Cell Line was generated using a lentiviral vector expressing the mouse-HVEM sequence.

Characterization

Figure 1: 293T-NFκB-Luc2-mouse-HVEM cell were seeded into 96-well plates, treated with Mouse LIGHT in different concentrations for 16 hours, and then read out using Bright-Glo Detection System.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS, 150μg/mL Hygromycin B and 1μg/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 1:4-1:6 every 2-3 days; seed out at about 1 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (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

1. Zhang T, Ye L, Han L, He Q, Zhu J. Knockdown of HVEM, a Lymphocyte Regulator Gene, in Ovarian Cancer Cells Increases Sensitivity to Activated T Cells. Oncol Res. 2016;24(3):189-96. doi: 10.3727/096504016X14641336229602. Erratum in: Oncol Res. 2017 Nov 2;25(9):1665. doi: 10.3727/096504017X15078984695565. PMID: 27458100; PMCID: PMC7838697.
2. Whitbeck JC, Connolly SA, Willis SH, Hou W, Krummenacher C, Ponce de Leon M, Lou H, Baribaud I, Eisenberg RJ, Cohen GH. Localization of the gD-binding region of the human herpes simplex virus receptor, HveA. J Virol. 2001 Jan;75(1):171-80. doi: 10.1128/JVI.75.1.171-180.2001. PMID: 11119586; PMCID: PMC113910.

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.