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Cat.No. : CSC-RR0552 Host Cell: SHP-77
| Cat. No. | CSC-RR0552 |
| Description | This cell line is engineered to stably express luciferase reporter gene in SHP-77. |
| Gene | Luciferase |
| Host Cell | SHP-77 |
| Host Cell Species | Homo sapiens (Human) |
| Stability | Validated for at least 10 passages |
| 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 |
Small cell lung cancer (SCLC) presents major clinical difficulties because of its aggressive character and poor prognosis. Though its exact function in SCLC is still unknown, recent investigations have shown MCM3AP-AS1, a long non-coding RNA (lncRNA), as an oncogene in many different malignancies. MCM3AP-AS1 is increased in SCLC patients, researchers observed, and associated with worse survival rates. Through qPCR, RNA pull-down, and luciferase assays, they demonstrated that MCM3AP-AS1 interacts with miR-148a, influencing ROCK1 expression, a gene associated with cancer cell invasion and migration. Overexpression of MCM3AP-AS1 promotes SCLC cell behaviors, while miR-148a counteracts these effects.
Figure 1. The researchers utilized RNA pull-down and luciferase assays, along with SHP-77 cells, to confirm the binding interaction between MCM3AP-AS1 and miR-148a. (Luo H, et al., 2021)
Creative Biogene's Luciferase Reporter Cell Line - SHP-77 is designed for similar investigations into gene regulation mechanisms in lung cancer and other malignancies.
A: The Luciferase Reporter Cell Line - SHP-77 can be employed to investigate the activation of specific signaling cascades by introducing it to various stimulators or inhibitors known to affect these pathways. Researchers can design experiments where the SHP-77 cells are treated with these agents and the resulting luciferase activity is measured over time using a luminometer or an imaging system capable of detecting bioluminescence.
A: To quantify the basal levels of luciferase activity in the SHP-77 cell line, researchers can use a luminescence assay, which typically involves lysing the cells and then measuring the light emitted upon the addition of a luciferin substrate.
Factors that might influence these measurements include cell density, passage number, and the health of the cells at the time of assay. It is important to maintain consistent culture conditions and to plate the cells at a uniform density to minimize variability. Additionally, researchers should ensure that the assay is performed under standardized conditions, such as using the same batch of luciferin substrate and following a consistent protocol for cell lysis and substrate addition.
A: The SHP-77 cell line offers several advantages for drug screening, including its ease of culture, its stable expression of the luciferase reporter, and its ability to respond to a wide range of pharmacological agents. The luciferase assay is highly sensitive and can be easily scaled up for high-throughput screening. To integrate the SHP-77 cell line into a high-throughput screening platform, researchers can automate the process of cell plating, treatment with test compounds, and luciferase activity measurement. This can be achieved using liquid handling robots, multi-well plate readers, and data analysis software capable of processing large datasets to identify compounds that modulate luciferase activity in a desired manner.
A: To validate the data obtained from the SHP-77 cell line, researchers can use complementary approaches such as quantitative PCR to measure the levels of the endogenous gene of interest, or Western blotting to detect the protein levels. Potential pitfalls in data interpretation include false positives due to non-specific effects of compounds on luciferase activity or cross-talk between different signaling pathways. To avoid these issues, researchers should perform dose-response experiments to confirm the specificity of the compound's effect and use multiple assays to confirm the biological activity of the compounds of interest. Additionally, it is important to consider the possibility of cytotoxic effects of the compounds, which can indirectly affect luciferase activity by altering cell viability.
A: When using the SHP-77 cell line to study the effects of environmental stressors, critical parameters that must be controlled include the duration and intensity of the stressor exposure, the timing of sample collection relative to stressor application, and the maintenance of a consistent culture environment (e.g., temperature, humidity, and CO2 levels). It is also important to include appropriate stressor controls, such as cells exposed to a non-stressful condition, to ensure that any changes in luciferase activity are specifically due to the environmental stressor. Researchers should also consider the potential for stressor-induced cytotoxicity, which can affect cell viability and confound the interpretation of luciferase activity data. By carefully controlling these parameters and using appropriate controls, researchers can accurately assess the impact of environmental stressors on gene expression as monitored by the luciferase reporter.
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The Luciferase Reporter Cell Line - SHP-77 is equipped with a luciferase reporter gene, making it highly sensitive for detecting cellular responses to stimuli. This sensitivity allows for the quantification of changes at very low levels, facilitating the study of subtle biological processes and improving the resolution of experimental outcomes.
Germany
04/12/2023
This cell line provides quantitative data outputs, allowing for precise measurement of gene expression levels. The Luciferase Reporter Cell Line - SHP-77's ability to emit light proportionally to the activity of certain promoters gives us exact numbers that are crucial for accurate interpretation and comparison of experimental results.
Germany
07/28/2020
The Luciferase Reporter Cell Line - SHP-77 offers rapid feedback on cellular responses due to the quick assay times associated with luciferase activity measurement. This rapid turnaround is essential for high-throughput screening and time-sensitive experiments, reducing the time from experiment to insight.
United States
06/07/2020
Using the Luciferase Reporter Cell Line - SHP-77 does not require cell destruction to measure reporter activity, allowing for longitudinal studies on the same cell populations. This non-destructive nature permits continuous monitoring of cellular responses over time, enhancing the understanding of dynamic biological processes.
Canada
02/07/2020
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