Cat.No. : CSC-RR0060 Host Cell: SK-OV-3
| Cat. No. | CSC-RR0060 |
| Introduction | The green fluorescent protein (GFP) is a protein composed of 238 amino acid residues (26.9kDa) that exhibits bright green fluorescence when exposed to light in the blue to ultraviolet range. Although many other marine organisms have similar green fluorescent proteins, GFP traditionally refers to the protein first isolated from the jellyfish Aequorea victoria. The GFP from A. victoria has a major excitation peak at a wavelength of 395 nm and a minor one at 475 nm. Its emission peak is at 509 nm, which is in the lower green portion of the visible spectrum. The fluorescence quantum yield (QY) of GFP is 0.79. The GFP from the sea pansy (Renilla reniformis) has a single major excitation peak at 498 nm. Luciferase is a generic term for the class of oxidative enzymes used in bioluminescence and is distinct from a photoprotein. One famous example is the firefly luciferase (EC 1.13.12.7) from the firefly Photinus pyralis. Firefly luciferase as a laboratory reagent usually refers to P. pyralis luciferase although recombinant luciferases from several other species of fireflies are also commercially available. |
| Gene | GFP/Luciferase |
| Host Cell | SK-OV-3 |
| Host Cell Species | Homo sapiens (Human) |
| Morphology | Epithelial |
| Stability | Validated for at least 10 passages |
| Reporter Type | Fluorescent protein |
| Application | 1. Gene expression studies 2. Protein localization 3. Drug screening and toxicology 4. Live cell imaging |
| Quality Control | Negative for bacteria, yeast, fungi and mycoplasma. |
| Shipping | Dry ice |
| Storage | Liquid nitrogen |
| Revival | Rapidly thaw cells in a 37°C water bath. Transfer contents into a tube containing pre-warmed media. Centrifuge cells and seed into a 25 cm2 flask containing pre-warmed media. |
| Growth Properties | Adherent |
| Mycoplasma | Negative |
| Format | One frozen vial containing millions of cells |
| Storage | Liquid nitrogen |
| Safety Considerations |
The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach. |
| Ship | Dry ice |
A: This cell line allows for the non-invasive real-time monitoring of gene expression by detecting the bioluminescent signal from luciferase and the fluorescent signal from GFP. These signals can be quantified using a luminometer and a fluorescence microscope or plate reader, respectively, to monitor gene expression dynamically in ovarian cancer studies.
A: The dual-reporter system enables concurrent monitoring of cell viability and reporter gene expression. GFP fluorescence can be used as a marker for cell health or transfection efficiency, while luciferase activity can indicate the activity of specific promoters or pathways targeted by drugs.
A: Yes, by treating these cells with anticancer drugs and measuring luciferase activity at various time points, researchers can determine the timing and duration of promoter activation or repression in response to treatment.
A: The cell line can be cultured under different microenvironmental conditions, such as hypoxia or in the presence of stromal cells. The effect of these conditions on gene expression can then be assessed by measuring GFP and luciferase signals, providing insights into how the tumor microenvironment influences ovarian cancer gene expression.
A: To quantify the effect of gene silencing, cells can be transfected with siRNAs targeting the gene of interest, followed by measurement of GFP and luciferase signals. A decrease in fluorescence and luminescence would indicate successful silencing and its consequent effect on promoter activity.
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The GFP/Luciferase Reporter Stable Cell Line-SKOV-3 utilizes both GFP (Green Fluorescent Protein) and luciferase as reporters, enabling both qualitative and quantitative analysis. This dual functionality allows us to visually track cell behavior with GFP while quantitatively measuring cellular responses via luciferase activity.
This cell line is engineered for high expression levels of both GFP and luciferase, ensuring that the fluorescent and luminescent signals are robust and easily detectable. High expression levels enhance the reliability and sensitivity of experimental assays involving these reporters.
GFP allows for real-time, non-destructive observation of cellular processes without the need for cell lysis. This is particularly valuable for dynamic studies where continuous monitoring is essential, as in the GFP/Luciferase Reporter Stable Cell Line-SKOV-3.
The stability of GFP and luciferase expression in the GFP/Luciferase Reporter Stable Cell Line-SKOV-3 ensures consistent performance over the course of long-term experiments. This stability is crucial for obtaining reliable, reproducible results in prolonged studies.
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