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luciferase Reporter Stable Cell Line-HEK293

luciferase Reporter Stable Cell Line-HEK293

Cat.No. :  CSC-RR0041 Host Cell:  HEK293

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Cat. No. CSC-RR0041
Description This cell line is engineered to stably overexpress the Luciferase in HEK293 cells.
Introduction Luciferase is a generic term for the class of oxidative enzymes used in bioluminescence and is distinct from a photoprotein. One famous example is the firefly luciferase (EC 1.13.12.7) from the firefly Photinus pyralis. Firefly luciferase as a laboratory reagent usually refers to P. pyralis luciferase although recombinant luciferases from several other species of fireflies are also commercially available.
Gene Luciferase
Host Cell HEK293
Host Cell Species Homo sapiens (Human)
Morphology Epithelial
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.
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
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Luciferase reporter stable cell line HEK293 is engineered to express a luciferase reporter gene for studying gene regulation and signal transduction pathways. The firefly-derived luciferase reporter gene produces an enzyme that catalyzes the oxidation of luciferin, which releases light. It is possible to quantify this emission in order to assess the activity of particular cell signaling pathways or promoters. Because of their strong proliferation and transfectability, human embryonic kidney cells, or HEK293 cells, are often utilized in scientific research. Since these cells are adherent, they may be grown under normal cell culture conditions with the right supplements and medium. They develop in a monolayer and have an epithelial shape, which makes them appropriate for experiments and high-throughput screening.

Progranulin plays a variety of roles in both physiological and pathological processes, such as neurodegeneration, inflammation, cancer, wound healing, and embryonic development. The HEK293 cells were used to create a stable cell line for the luciferase reporter to study the transcriptional regulation of the PGRN gene. The luciferase gene was integrated into the genome using CRISPR/Cas9, with the native PGRN promoter serving as its master regulator. The luciferase gene's site-specific integration was validated by PCR findings. Applying a CRISPR/Cas9 transcription activation/repression system and treating the cells with phorbol ester (phorbol 12-myristate, 13-acetate), which is known to activate PGRN expression, allowed them to validate this approach. The direct correlation between the luciferase activity and the PGRN promoter activity suggests that the approach is useful for investigating the transcriptional regulation of PGRN and may have applications in drug screening that targets PGRN.

Figure 1 depicts the regulation of the endogenous promoter of the PGRN gene by the CRISPR/Cas9 system. The transcriptional activation effects of different single sgRNAs (a) and multiple sgRNAs (b) on the exogenous PGRN promoter activity in HEK293 cells were compared. (doi: 10.1080/21655979.2019.1607126)Figure 1. Using CRISPR/Cas9 systems, the researchers screened the effectiveness of sgRNA in controlling hPGRN gene expression using the Luciferase Reporter Stable Cell Line-HEK293; luciferase activity and real-time PCR analysis were then employed. (Li Y, et al., 2019)

1. Gene Regulation Research: Using the luciferase Reporter stable cell line HEK293, we studied the gene regulation mechanism, especially explored the role of promoter activity and transcription factors, so as to gain a deeper understanding of the gene expression regulatory network. 2. Signal Transduction Pathway: Using the HEK293 cell model, we studied the activity of different signal transduction pathways, and used the luciferase reporter gene to detect the level of intracellular active molecules to promote the analysis of cell signal transduction mechanisms. 3. Drug Screening and Evaluation: Using the luciferase Reporter cell line, we screened and evaluated drugs, evaluated the effects of potential drugs on specific signal pathways, and accelerated the process of new drug development. 4. Cell Proliferation and Transfection: With the strong proliferation ability and easy transfection of HEK293 cells, we conducted experiments such as gene editing and gene transduction to explore the mechanisms of cell growth, proliferation and transfection. 5. High-throughput Screening: Using the epithelial morphology and easy-to-culture characteristics of HEK293 cells, we conducted high-throughput screening experiments to quickly screen out cell lines with specific functions, providing tools and methods for functional genomics research.
Customer Q&As
What is the Luciferase expression level of Luciferase Reporter Stable Cell Line-HEK293 cell line? Are there fluctuations in gene expression levels?

A: Our Luciferase Reporter cell lines have been rigorously screened and verified to ensure stable expression of Luciferase. We verified the expression level of Luciferase through methods such as luciferase activity detection and PCR, and repeated tests were performed at different time points and in different batches. Cells express Luciferase under appropriate culture conditions and maintain stable expression levels.

How does the HEK293 background of this Reporter cell line affect the performance of the product? Are there any special considerations related to this cell line?

A: Our Luciferase Reporter cell line has demonstrated excellent performance in the HEK293 cell line. During use, we recommend that customers pay close attention to the growth of cells and follow the culture guidelines we provide. Our cell lines have been optimized to ensure optimal expression and stability in the HEK293 cell line.

Has the HEK293 cell line been screened and optimized for this product? How does the screening process work?

A: Yes, our HEK293 cell line has undergone a rigorous screening and optimization process. We use specific screening criteria, including luciferase activity assays and gene expression levels, to ensure that the stability and expression levels of the cell lines meet our quality standards. We also performed long-term culture and expansion of the cell line to verify its stability and reproducibility under different conditions.

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Customer Reviews
Stability

I purchased Luciferase Reporter Stable Cell Line-HEK293 using this supplier's platform and I am very pleased with the growth of the cells. During the culture process, I observed very consistent cell growth with no abnormalities. Gene expression is also very stable, in line with my expectations. I can easily assess the activity of cell signaling pathways, which is very helpful in my research. The products provided by the supplier did meet my expectations and I was very impressed with their service.

United Kingdom

07/16/2022

growth consistency

I am very satisfied with the Luciferase Reporter Stable Cell Line-HEK293 provided by this platform. After I purchased them, I found that the cells grew very stably and had very consistent gene expression. This makes my experimental work smoother and more efficient. I particularly like the easy transfection nature of these cells and their adaptability under normal cell culture conditions. This is indeed a high quality cell line and I will continue to choose products from this supplier.

United Kingdom

01/13/2020

Easy to transfect

Gene expression is also very stable, in line with my expectations. Easy to operate, they have great applicability in experiments and high-throughput screening. This purchasing experience left me deeply impressed with the products of this supplier, and I will continue to pay attention to their new products.

Germany

08/06/2022

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