KC-6346

LNCap-GFP-Luc2 Cell-Line

66236

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Background of LNCap-GFP-Luc2 Cell-Line

Luciferase is an oxidative enzyme that can produce bioluminescence with addition of luciferin, but doesn’t need an external light source unlike fluorescent proteins. Photo emission can be detected directly by light sensitive devices. Such as luminometers or modified microscopes. Luciferase is widely used in many fields of biological research, such as transcriptional activity, kinase or other enzyme activity, cellular ATP level, and whole animal imaging.
Green fluorescent protein (GFP) is a β - barrel protein 1 composed of 238 amino acids with a molecular weight of approximately 27 kDa. GFP was isolated from the crystal jellyfish Aequorea Victoria. GFP can convert the blue fluorescence emitted by jellyfish luminescent protein through chemical reactions into green fluorescence through energy transfer. The excitation wavelength of GFP is 488 nm, and there is an emission peak at approximately 507 nm.

Specifications

Catalog Number	KC-6346
Cell Line Name	LNCap-GFP-Luc2 Cell-Line
NCBI/UniProt Accession Number	N/A
Clone Number	3#
Host Cell Line	LNCap cell line
Description	Stable LNCap cell line expressing exogenous GFP and Luc2 reporter genes.
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% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS
Selection Marker	N/A
Morphology	Fibroblast-like
Subculture	Split saturated culture 1:2-1:4 every 3-4 days
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative
In Vivo Validation	N/A

Cell Line Generation

The LNCap-GFP-Luc2 cell line was generated using a lentiviral vector expressing the GFP and Luc2 sequences.

Characterization

Cell Resuscitation

Pre-warm complete culture medium (basal medium and 10% FBS) in a 37°C water bath.
Rapidly thaw the cryovial in a 37°C water bath for 1-2 minutes with gentle agitation.
Transfer the vial to a biosafety cabinet, and disinfect the exterior with 70% ethanol.
Aseptically transfer the cell suspension dropwise into a sterile centrifuge tube containing 9.0 mL of pre-warmed complete medium.
Centrifuge at approximately 125 × g for 5–7 minutes at room temperature, carefully aspirate the supernatant without disturbing the cell pellet.
Gently resuspend the pellet in an appropriate volume of complete medium and transfer the suspension into a T25 flask.
Incubate the flask in a 37°C in a humidified 5% CO₂ incubator.
Assess cell viability and morphology after 24 hours. If cells appear healthy, replace the medium with fresh medium supplemented with the appropriate selective antibiotic.
Subculture the cells at a ratio of 1:2-1:4 every 3-4 days upon reaching 80%–90% confluency.

Cell Freezing

Prepare the freezing medium (70% basal medium, 20% FBS and 10% DMSO) freshly before use.
Pre-chill the freezing medium on ice and label the cryovials accordingly.
Transfer the cell suspension to a sterile conical tube and perform a cell count to determine total viability and density.
Centrifuge the cells at 250×g for 5 minutes at room temperature; carefully aspirate the supernatant.
Gently resuspend the cell pellet in chilled freezing medium, ensuring a minimum cell density of 3×10⁶ cells/mL.
Aliquot 1 mL of the cell suspension into each pre-labeled cryovial.
Place the cryovials into a CoolCell® container and store at -80°C overnight for controlled-rate cooling.
Transfer the cryovials to the liquid nitrogen for long-term storage the following day.

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.
2.Pieribone V, Gruber D. Aglow in the Dark: The Revolutionary Science of Biofluorescence. Cambridge: Belknap Press. 2006. ISBN 0-674-01921-0. OCLC 60321612. Popular science book describing history and discovery of GFP

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.