Cat.No. : CSC-RR0523 Host Cell: MC38
| Cat. No. | CSC-RR0523 |
| Description | This cell line is engineered to stably overexpress ZsYellow1 reporter gene. MC38 is a murine colon carcinoma cell line, which has been widely used in laboratory research. This cell line is a useful tool for fluorescent tracking of MC38 cells. |
| Gene | ZsYellow1 |
| Host Cell | MC38 |
| Host Cell Species | Mus musculus (Mouse) |
| 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 |
A: When optimizing culture conditions for the ZsYellow1 Reporter Cell Line - MC38, researchers should consider factors such as the appropriate cell growth medium, which typically includes a balanced mixture of nutrients, growth factors, and hormones; the temperature and CO2 concentration in the incubator to maintain the cells' physiological state; and the passage number to avoid senescence, which can affect protein expression levels. Additionally, regular monitoring of cell density and subculturing practices are crucial to maintain the health and viability of the cells, which in turn affects the reliability of the ZsYellow1 fluorescence as a reporter.
A: The ZsYellow1 fluorescence intensity can be quantitatively correlated with specific biological processes by first validating the response of the reporter to the process under study. This typically involves treating the cells with known inducers or inhibitors of the target process and measuring changes in fluorescence over time using techniques such as flow cytometry or fluorescence microscopy.
Researchers must establish a standard curve by treating the cells with varying concentrations of the inducer and measuring the resulting fluorescence. This curve can then be used to estimate the activity of the biological process in unknown samples based on their fluorescence intensity. It is essential to normalize the fluorescence data against an internal control, such as cell count or protein content, to account for variability in cell number or health.
A: Potential limitations of using the ZsYellow1 Reporter Cell Line - MC38 in high-throughput screening include the possibility of non-specific fluorescence from other cellular components, photobleaching of the ZsYellow1 protein under prolonged illumination, and the potential for the reporter to be affected by factors unrelated to the target process.
To mitigate these limitations, researchers can use appropriate controls, such as a non-reporter cell line or cells with a different reporter, to account for background fluorescence. Additionally, optimizing the imaging or measurement conditions to minimize light exposure and using protective measures like anti-fade reagents can help prevent photobleaching. Finally, careful experimental design and statistical analysis can help distinguish true signals from noise and account for confounding factors.
A: The ZsYellow1 Reporter Cell Line - MC38 is specifically engineered to express the ZsYellow1 protein, which offers a high signal-to-noise ratio and is spectrally distinct from other commonly used fluorescent proteins. This allows for the specific detection of ZsYellow1 fluorescence without interference from other cellular fluorophores.
The MC38 cell line, being derived from a colon carcinoma, provides a relevant model for studying cancer-related processes. The advantages of using this particular cell line include its well-characterized growth properties and its amenability to genetic manipulation, which can facilitate the study of complex cellular pathways. Moreover, the use of a single reporter system within the cell line simplifies data interpretation compared to multi-reporter systems, where cross-talk between reporters can be an issue.
A: To validate the specificity of the ZsYellow1 fluorescence signal in the MC38 cell line, researchers must first confirm that the fluorescence is indeed due to the expression of the ZsYellow1 protein by using techniques such as Western blotting or immunofluorescence staining.
Additionally, researchers should demonstrate that the fluorescence signal is modulated only by the target process and not by other cellular changes. This can be achieved by comparing the ZsYellow1 signal with an independent measure of the target process or by using RNA interference or CRISPR/Cas9 gene editing to specifically knock down or knockout the gene of interest and observing the effect on the fluorescence signal.
Furthermore, it is important to rule out any non-specific effects on fluorescence, such as changes in cell morphology or autofluorescence from other cellular components, by using appropriate controls and imaging settings. By taking these critical steps, researchers can ensure that the observed changes in ZsYellow1 fluorescence accurately reflect the biological process under investigation.
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ZsYellow1 Reporter Cell Line - MC38 expresses bright yellow fluorescence, making cells easily distinguishable under fluorescence microscopy, which aids in cell tracking and localization in complex tissues or co-culture conditions.
The yellow fluorescence of ZsYellow1 in the ZsYellow1 Reporter Cell Line - MC38 penetrates deeper into tissues compared to other fluorescence colors, improving visibility in in vivo imaging applications.
This cell line ensures stable expression of ZsYellow1, providing consistent and reliable fluorescent signals across various experimental conditions and over extended culture periods.
ZsYellow1 offers high contrast against common lab stains and background fluorescence, which simplifies analysis and increases accuracy in quantifying cell behavior and population dynamics in studies using the ZsYellow1 Reporter Cell Line - MC38.
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