Cat.No. : CSC-SC015138-1 Host Cell: HEK293
| Cat. No. | CSC-SC015138-1 |
| Description | This cell line is engineered to stably express human steroid 5 alpha-reductase 1 (SRD5A1) in HEK293 cells. |
| Gene | SRD5A1 |
| Gene Species | Homo sapiens (Human) |
| Host Cell | HEK293 |
| Host Cell Species | Homo sapiens (Human) |
| Stability | Validated for at least 10 passages |
| Application | 1. Gene expression studies 2. Signaling pathway research 3. Drug screening and toxicology 4. Disease research |
| 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 |
The enzyme steroid 5-α reductase (5AR) is in charge of converting steroids into 5-α reduced metabolites, like 5-α dihydrotestosterone (DHT) from testosterone. To evaluate the inhibition of steroid 5-α reductase (5AR), which is essential for converting steroids like testosterone to 5-α dihydrotestosterone (DHT), the researchers created a cell-based assay. They determined the IC50 values for the 5AR inhibitors finasteride and dutasteride using human and fish cell lines in conjunction with a transient overexpression technique. It's interesting to note that fish cell lines had a lower IC50 for dutasteride than human cell lines, suggesting species-specific responses. This sensitive test exhibits excellent reproducibility and is appropriate for screening endocrine-disrupting chemicals (EDCs). It makes use of chemical derivatization and liquid chromatography-tandem mass spectrometry. Subsequent investigations will delve deeper into the inhibition of 5AR by EDCs in other species to clarify their influence on endocrine pathways.
Figure 1. The researchers used non-vector- and SRD5A1-HEK293 cells and SRD5A2-HEK293 cells to measure DHT levels after testosterone addition. KM values were calculated based on the enzyme's affinity. Inhibition assays with finasteride and dutasteride followed, determining IC50 values. (Kim D, et al., 2021)
A: Human SRD5A1 Stable Cell Line - HEK293 is specifically engineered for stable SRD5A1 expression in HEK293 cells. Its stability is rigorously tested over long passages to ensure consistent and reliable expression levels.
A: Authentication of SRD5A1 expression in Human SRD5A1 Stable Cell Line - HEK293 involves techniques such as PCR sequencing, Western blotting, and immunofluorescence staining. These methods confirm the presence and integrity of the SRD5A1 gene and protein.
A: Yes, Human SRD5A1 Stable Cell Line - HEK293 is suitable for functional assays assessing SRD5A1 activity, such as enzymatic assays measuring substrate conversion rates. These assays provide valuable insights into the functionality of expressed SRD5A1 protein.
A: Off-target effects in Human SRD5A1 Stable Cell Line - HEK293 are minimized through careful selection of stable integration sites and thorough screening for single-copy integration. Additionally, control experiments with wild-type cells serve to validate specific effects.
A: Strategies to enhance SRD5A1 expression in Human SRD5A1 Stable Cell Line - HEK293 include optimizing promoter sequences, codon usage, and post-translational modification pathways. These optimizations improve protein yield and stability.
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HEK293 cells are stably engineered to ensure long-term stable expression of SRD5A1.
Human SRD5A1 Stable Cell Line generates a large quantity of SRD5A1, providing ample samples for research.
The cell line is easy to culture and handle, providing convenience to researchers.
Each batch of Human SRD5A1 Stable Cell Line maintains consistent performance and expression levels, ensuring result reproducibility.
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