Cat.No. : CSC-RR0529 Host Cell: A20
| Cat. No. | CSC-RR0529 |
| Description | This cell line is engineered to stably overexpress mCherry reporter gene. A20 is a murine lymphoma cell line, which has been widely used in laboratory research. This cell line is a useful tool for fluorescent tracking of A20 cells. |
| Gene | mCherry |
| Host Cell | A20 |
| 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: For quantifying mCherry fluorescence intensity in the mCherry Stable Cell Line - A20, flow cytometry and fluorescence microscopy are the most effective methodologies. Flow cytometry allows for the rapid quantification of fluorescence in thousands of cells per sample, providing a robust statistical analysis of mCherry expression levels across a cell population. Fluorescence microscopy, on the other hand, enables spatial resolution of mCherry expression within individual cells, allowing researchers to observe localization and distribution patterns. The choice of methodology can impact experimental outcomes by affecting the type of data collected (quantitative population data vs. qualitative single-cell data), and thus, should be aligned with the specific objectives of the research project.
A: The mCherry Stable Cell Line - A20 can be utilized to study protein-protein interactions within the NF-kB signaling pathway by creating fusion proteins of mCherry with various components of the NF-kB pathway. These fusion constructs can be transfected into the A20 cells, and the localization, movement, and interaction of these proteins can be monitored in real-time using live-cell fluorescence microscopy. This approach allows researchers to visually track the dynamics of NF-kB signaling components in the context of cellular responses to stimuli, providing insights into the spatial and temporal aspects of protein-protein interactions within this critical signaling pathway.
A: Challenges associated with using the mCherry Stable Cell Line - A20 for live-cell imaging include photobleaching of the mCherry fluorophore, potential phototoxicity to cells, and maintaining cell health under prolonged imaging conditions. To overcome these challenges, researchers can minimize exposure to intense light by using low-light imaging techniques and employing filters that specifically match the mCherry excitation/emission wavelengths to reduce photobleaching. Additionally, optimizing the imaging medium to support cell health and using temperature and CO2 control systems for live-cell chambers can help maintain cell viability during extended observations.
A: The expression of mCherry in the A20 cell line facilitates the study of cell cycle dynamics and cellular proliferation by providing a fluorescent marker that can be continuously monitored in live cells. By coupling mCherry expression with cell cycle-specific markers or using time-lapse fluorescence microscopy, researchers can track the progression of individual cells through the cell cycle. This approach allows for the direct observation of mitotic events, changes in cell morphology, and the dynamics of cell division. Furthermore, analyzing changes in mCherry fluorescence intensity over time can help quantify rates of cellular proliferation, enabling studies on how various factors influence cell cycle progression in the A20 cell line.
A: The use of the mCherry Stable Cell Line - A20 can contribute to the development of gene therapy vectors by serving as a reporter system to evaluate the efficiency of vector-mediated gene delivery. The mCherry expression provides a readily quantifiable measure of transduction efficiency, allowing researchers to optimize vector constructs, delivery methods, and transgene expression levels. When employing this cell line for vector testing, considerations should include ensuring the compatibility of the vector with the A20 cell line, determining the optimal multiplicity of infection (MOI) for efficient transduction without cytotoxicity, and validating that the vector does not interfere with the endogenous mCherry expression. Additionally, comparing the performance of different vector designs or delivery methods can be facilitated by quantifying mCherry fluorescence as a surrogate for successful gene delivery.
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The mCherry Stable Cell Line - A20 utilizes mCherry, a bright red fluorescent protein, which offers excellent visibility under fluorescence microscopy. This bright tagging allows for clear and distinct visualization of cell structures and dynamics, making it easier to conduct detailed cellular studies without the interference from autofluorescence often present in biological samples.
Engineered to stably express mCherry, the mCherry Stable Cell Line - A20 ensures consistent fluorescence across different experimental conditions and over extended culture periods. This stable expression is crucial for long-term studies, where consistent protein expression is necessary to obtain reliable and reproducible data.
mCherry in the mCherry Stable Cell Line - A20 allows for non-invasive live-cell monitoring. We can observe and record cellular processes in real-time without the need for cell fixation or staining, preserving the natural state of cells and providing more accurate insights into cellular behaviors.
The distinct red fluorescence of mCherry makes the mCherry Stable Cell Line - A20 ideal for co-localization studies with other fluorescently tagged proteins. This capability is essential for studying protein-protein interactions within the cell, facilitating a deeper understanding of cellular mechanisms and signaling pathways.
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