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Cat.No. : CSC-RR0280 Host Cell: SK-BR-3
| Cat. No. | CSC-RR0280 |
| Description | SKBR3 - GFP cell line has been developed through stable transfection with GFP. GFP is expressed as free cytoplasmatic proteins in SKBR3 - GFP cells. This cell line is a ready-to-use in vitro model for cell-based assay applications. |
| Gene | GFP |
| Host Cell | SK-BR-3 |
| 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. |
| Storage | Liquid nitrogen |
| Source | SKBR3 |
| 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 |
Researchers used a combination of SERS and Raman technology to identify and accurately evaluate HER2-expressing cells. The researchers selected the SKBR3 cell line with the highest HER2 expression level, the MDA-MB-468 cell line with the lowest level, and the normal breast epithelial cell line MCF10A for comparison. To further verify the effectiveness of this method, the researchers used RNA interference technology to silence the HER2 gene in SKBR3 cells and generate stable clonal cell lines. Using this combined approach, they analyzed cells before and after HER2 silencing, detected significant changes in HER2 expression at the single-cell level, and observed that HER2 status correlated with changes in fatty acid content and downstream signaling molecule content.
Figure 1. The researchers used the GFP Reporter Cell Line - SKBR3 cell line to simulate the effects of HER2-targeted therapy. They stably silenced the HER2 gene by RNA interference technology and analyzed the obtained clones using the SERS-Raman method. The researchers transfected SKBR3 cells with a plasmid containing a puromycin resistance marker, a GFP encoding gene, and a shRNA transcription unit targeting HER2 mRNA. Using methods such as fluorescence microscopy, Western blot, and flow cytometry, they evaluated the transfection efficiency and changes in HER2 expression levels and selected clones with the best silencing effect for subsequent studies. (Spaziani S, et al., 2024)
A: Researchers can use the GFP Reporter Cell Line - SKBR3 to evaluate the impact of anti-cancer compounds on cell cycle regulation and apoptosis by monitoring changes in GFP fluorescence, which correlates with cellular processes. By treating the cells with various concentrations of the test compounds and analyzing the GFP expression levels and patterns, one can assess the effects on cell cycle progression and the induction of apoptosis. This approach allows for the identification of compounds that effectively modulate these processes in breast cancer cells, potentially leading to the development of novel therapeutic strategies.
A: When employing the GFP Reporter Cell Line - SKBR3 for high-throughput screening, it is crucial to standardize experimental conditions such as cell seeding density, incubation time, and media composition to ensure consistent GFP expression. Additionally, the choice of appropriate fluorescence detection equipment and data analysis software is essential for accurate quantification of GFP signal. Researchers must also consider the use of appropriate controls, including positive and negative controls, to validate the screening results.
A: By introducing pathway-specific inhibitors or activators into the culture of the GFP Reporter Cell Line - SKBR3, researchers can study the influence of these pathways on the migration and invasion capabilities of breast cancer cells. The GFP fluorescence allows for the tracking of cellular behavior, providing a visual readout of the effects on cell motility. This can help elucidate the molecular mechanisms underlying breast cancer metastasis and identify potential therapeutic targets.
A: The GFP Reporter Cell Line - SKBR3 offers a unique opportunity to study the impact of environmental stressors, such as hypoxia or oxidative stress, on breast cancer cells. The GFP reporter serves as a sensitive indicator of changes in cellular stress responses, allowing researchers to monitor alterations in cell behavior and survival under stress conditions. This can provide insights into the adaptive mechanisms of breast cancer cells and contribute to the understanding of tumor resilience and progression.
A: The GFP Reporter Cell Line - SKBR3 can be used to model the response of individual patients to targeted therapies by introducing patient-derived genetic variations into the cell line. By correlating GFP expression patterns with the presence or absence of specific genetic markers, researchers can predict the efficacy of targeted drugs and contribute to the development of personalized treatment strategies for breast cancer patients.
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By linking GFP expression to gene promoters, the GFP Reporter Cell Line - SKBR3 becomes a valuable tool for studying promoter activity and gene regulation in breast cancer cells.
United States
03/12/2020
Equipped with GFP, the cell line simplifies various cellular assays, as no additional staining is needed to visualize SKBR3 cells under fluorescence microscopy.
Germany
10/14/2022
The GFP Reporter Cell Line - SKBR3 enables us to easily assess transfection efficiency, streamlining gene expression studies and molecular assays.
Canada
03/17/2022
With stable GFP integration, this cell line ensures consistent fluorescence, critical for comparative studies and long-term observation in GFP Reporter Cell Line - SKBR3.
Germany
08/04/2023
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