Cat.No. : CSC-RR0066 Host Cell: HEK293
| Cat. No. | CSC-RR0066 |
| Description | This cell line is engineered to stably overexpress the RFP in HEK293 cells. |
| Background | The development of the RFP Reporter Stable Cell Line-HEK293 traces its origins to the early 1970s with the discovery and establishment of the Human Embryonic Kidney 293 (HEK293) cell line by Graham et al. in 1977. Initially derived from human embryonic kidney cells |
| Introduction | Red Fluorescent Protein (RFP) is a genetic mutant of green fluorescent protein. |
| Gene | RFP |
| Host Cell | HEK293 |
| Host Cell Species | Homo sapiens (Human) |
| Morphology | Epithelial |
| 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. |
| Media Type | Cells were cultured in DMEM supplemented with 10% fetal bovine serum. |
| Freeze Medium | Complete medium supplemented with 10% (v/v) DMSO |
| 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. |
| Growth Properties | Cells are cultured as a monolayer at 37°C in a humidified atmosphere with 5% CO2. Split at 80-90% confluence, approximately 1:3-1:6. |
| 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 |
Posttranslational modifications extensively alter the ubiquitous transcription factor, specificity protein 1 (SP1). Researchers employ RFP Reporter Stable Cell Line-HEK293 to probe SP1's role in Wnt signaling. This cell line facilitates studying SP1's stability modulation by Wnt components like GSK3β, β-TrCP E3 ubiquitin ligase, and β-catenin. Monitoring RFP reporter expression allows dynamic assessment of SP1 degradation upon Wnt activation, unveiling SP1-β-catenin crosstalk's impact on target gene expression. This study unveils pivotal insights into SP1-β-catenin interplay in Wnt signaling.
Figure 1. Researchers discovered that RFP SP1 is vital for stabilizing β-catenin and regulating Wnt-responsive genes. Immunofluorescence analysis in siSP1-transfected HCT116 cells showed reduced nuclear SP1 and β-catenin. Immunoblots in Wnt3A-treated HEK293 cells upon SP1 depletion exhibited altered SP1, β-catenin, and TCF7 levels. (Mir R, et al., 2018)
A: HEK293 cells were chosen for establishing the RFP reporter stable cell line due to their similar advantages to the GFP reporter line, including high transfection efficiency and suitability for protein expression studies. Additionally, RFP provides an alternative fluorescence marker for multiplexing experiments.
A: The stability and expression level of RFP in the HEK293 stable cell line was verified and sustained through stable transfection and antibiotic selection. Expression was validated using fluorescence microscopy or flow cytometry, and stability was ensured through regular culture maintenance and monitoring of fluorescence intensity.
A: Insights into the functional characterization of RFP expression in the HEK293 stable cell line included assessing its responsiveness to regulatory elements through changes in fluorescence intensity in response to stimuli. Additionally, RFP's involvement in cellular processes such as protein trafficking or organelle labeling was investigated using live-cell imaging or co-localization studies.
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Consistent expression! This cell line surpasses expectations, providing bright and uniform RFP fluorescence, facilitating accurate quantification and visualization of cellular processes. Its stable RFP expression simplifies experimental workflows, allowing for rapid data acquisition and analysis, and accelerating discoveries in cell biology and molecular biology.
Remarkable fluorescence! The RFP Reporter Stable Cell Line in HEK293 cells offers stable expression of RFP, enabling easy visualization and tracking of cellular events in my research.
The RFP Reporter Stable Cell Line has significantly enhanced my research capabilities, offering a versatile platform for studying gene expression dynamics and protein localization in vitro.
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