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
Price:
0
Host Cell Line:
Human A431 cell line
Description:
Stable A431 clone expressing exogenous luciferase
Quantity:
One vial of frozen cells (5X106 per vial)
Stability:
Stable in culture over a minimum of 10 passages
Application:
Drug screening and biological assays
Freeze Medium:
70% DMEM+20% FBS+10% DMSO
Propagation Medium:
DMEM+10%FBS+100µg/mL Hygromycin B
Selection Marker:
HygromycinB
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
Cell Line Generation
A431 Luc2 cell line was generated using a lentiviral vector expressing luciferase sequence.
Characterization using PCR sequencing and WB
0
Application
0
Cell Resuscitation
1. Prewarm culture medium (DMEM + 10% FBS + 100µg/mL Hygromycin B) in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO2 incubator.
8. Split saturated culture 1:2 every 3-5 days; seed out at about 1-3 × 105 cells/mL.
Cell Freezing
1. Prepare the freezing medium (70% DMEM + 20% FBS + 10% DMSO) fresh immediately before use.
2. Keep the freezing medium on ice and label cryovials.
3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3×106 cells/mL in chilled freezing medium.
6. Aliquot 1 mL of the cell suspension into each cryovial.
7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
8. Transfer vials to liquid nitrogen for long-term storage
References
1. Greer LF, Szalay AA (2002). "Imaging of light emission from the expression of luciferases in living cells and organisms: a review". Luminescence. 17 (1): 43–74. doi:10.1002/bio.676. PMID 11816060.
