KC-6138

22RV1-CHD7-KO-mcherry-Luc2 Cell Line

60818

Home » 22RV1-CHD7-KO-mcherry-Luc2 Cell Line

Background of 22RV1-CHD7-KO-mcherry-Luc2 Cell Line

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

Specifications

Catalog Number	KC-6138
Cell Line Name	22RV1-CHD7-KO-mcherry-Luc2 Cell Line
NCBI/UniProt Accession Number	N/A
Clone Number	14#
Host Cell Line	22RV1-CHD7-KO
Description	Stable 22RV1-CHD7-KO clone with mcherry and 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	RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10% FBS+300μg/mL Hygromycin B
Selection Marker	Hygromycin B
Morphology	Epithelial
Subculture	Split saturated culture 1:2 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

Cell Line Generation

22RV1-CHD7-KO-mcherry-Luc2 Cell Line was generated using a lentiviral vector expressing mcherry and luciferase sequence.

Characterization

Figure 1: Characterization of 22Rv1-CHD7-KO cell line stable clone using PCR sequencing

Figure 2: Characterization of mcherry overexpression in the 22RV1-CHD7-KO-mcherry-Luc2 stable clone using FACS.

Figure 3: Characterization of luciferase expression in 22RV1-CHD7-KO-mcherry-Luc2 cell using the Bright-Glo Detection.

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS + 300μg/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 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

Greer LF 3rd,Szalay AA.Imaging of light emission from the expression of luciferases in living cells and organisms:a review.Luminescence.2002 Jan-Feb;17(1):43-74.doi:10.1002/bio.676.PMID:11816060.