KC-2592

A431-Luc2-Cell-Line

24366

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Background of A431-Luc2-Cell-Line

Luciferase is an oxidative enzyme that can produce bioluminescence with addition of luciferin, but don’t need an external light source unlike of fluorescent proteins. Photo emission can be detected directly by light sensitive device. Such as luminometer or modified microscope. Luciferase is widely used in many fields of biological research, such as transcriptional activity, kinase or other enzyme activity, cellular ATP level, whole animal imaging.

Specifications

Catalog Number	KC-2592
Cell Line Name	A431-Luc2-Cell-Line
Host Cell Line	Human A431 cell line
Description	A431 cell line stable clone with Luciferase gene overexpression
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+100µg/mL Hygromycin B
Selection Marker	Hygromycin B
Morphology	Epithelial
Subculture	Split saturated culture 1:2 every 3-5 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 80-100 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

A431 Luc2 cell line was generated using a lentiviral vector expressing luciferase sequence.

Characterization

Figure : Characterization of luciferase expression in A431 using the Bright-Glo Detection.

Cell Resuscitation

Prewarm culture medium (DMEM + 10% FBS + 100µg/mL Hygromycin B) 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 1:2 every 3-5 days; seed out at about 1-3 × 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

Greer LF, Szalay AA (2002). "Imaging of light emission from the expression of luciferases in living cells and organisms: a review". Luminescence. 17 (1): 43ÿ74. doi:10.1002/bio.676. PMID 11816060.