Cat.No. : CSC-RR0436 Host Cell: HCT116
| Cat. No. | CSC-RR0436 |
| Description | HCT116-GFP/RFP cell line is engineered to stably express H2B-EGFP and RFP. It is a good ready-to-use cell model for tracing HCT116 cells. |
| Host Cell | HCT116 |
| Host Cell Species | Homo sapiens (Human) |
| 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. |
| 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 |
The GFP/RFP Reporter Cell Line-HCT116 was used by the researchers to investigate the function of NRF3 in cancer cells. They found that NRF3 increases the viability of cancer cells by activating mTORC1 in response to amino acids, particularly arginine. NRF3 also upregulates SLC38A9 and RagC expression, which facilitates mTORC1 recruitment to lysosomes. Furthermore, NRF3 promotes RAB5-mediated macropinocytosis and SLC7A1-mediated arginine transport into lysosomes. Finally, NRF3 promotes RAB5-mediated macropinocytosis and SLC7A1-mediated arginine transport into lysosomes. Finally, inhibiting the NRF3–mTORC1 axis impairs mitochondrial function, which leads to cancer cell apoptosis.
Figure 1. The GFP/RFP Reporter Cell Line-HCT116 was used by the researchers to evaluate the effect of NRF3 on mTORC1 activity. They used GFP/RFP fluorescence ratios to measure autophagy flux and carried out siRNA transfections. (Hirose S, et al., 2023)
A: The HCT116 GFP/RFP reporter cell line exhibits excellent fluorescence intensity and signal stability by stably expressing green fluorescent protein (GFP) and red fluorescent protein (RFP). The fluorescence signal of this cell line remains stable after multiple passages and can provide consistent fluorescence signals under different experimental conditions. This stability helps reduce signal fluctuations during long experiments, ensuring data reliability and repeatability. In addition, the cell lines undergo strict quality control and validation, ensuring their high quality and experimental reproducibility.
A: When using the HCT116 GFP/RFP reporter cell line to perform dual fluorescence reporter experiments, special attention should be paid to the resolution and detection of fluorescent signals. First, select appropriate fluorescence filters and microscope settings to ensure that GFP and RFP signals can be clearly distinguished and avoid cross-interference. Secondly, cell culture conditions should be consistent, including temperature, CO2 concentration and culture medium components, to ensure normal cell growth and expression of fluorescent proteins. In addition, before performing fluorescence imaging or fluorescence intensity measurement, the cell density should be ensured to be moderate to prevent excessive confluence of cells from affecting the accuracy of the fluorescence signal.
A: To ensure that the fluorescence data generated by the HCT116 GFP/RFP reporter cell line is accurate and consistent, it is recommended to take the following measures: First, repeat the experiment multiple times to obtain statistically significant data, thereby reducing the impact of accidental errors. Secondly, use standardized fluorescence measurement methods and equipment to ensure data comparability. At the same time, during the data analysis process, appropriate software should be used to perform quantitative analysis of fluorescence intensity and correct for background fluorescence. In addition, record all variables of the experiment, including cell culture conditions, fluorescence measurement parameters, etc., for comparison and verification in subsequent experiments.
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The GFP/RFP reporter cell line provided by this supplier is quite good. During the culture process, the growth of cells is relatively stable, and gene expression is relatively consistent. Sometimes you will see some changes in the intensity of red and green fluorescence, but overall, you can still observe the dynamics of cells relatively clearly. However, the cell proliferation rate is slightly faster and requires more attention.
The GFP/RFP reporter cell line of this platform has been used and the results are good. The cells grew very fast during the culture process and the fluorescence signal was stable. Overall, the dynamics of cells can be tracked and the data is accurate. In addition, the morphology of the cells was maintained well, and they were basically in the form of epithelial cells.
The performance of this company's GFP/RFP reporter cell line is quite satisfactory. During the experiment, the cells proliferated rapidly, in line with expectations. The morphology of the cells is okay, they are basically epithelial cells.
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