商品编号


SL-0011-FP



品牌


Signosis



公司


Signosis



公司分类


Cell-Based Assays




1 Ea


商品信息

Description:
p53 Responsive Luciferase Reporter Hela Stable Cell Line is derived from human cervical cancer? , and stably express firefly luciferase reporter gene under the control of the p53 response element.? This cell line is an ideal cellular model for monitoring the activation of p53 Receptor Signaling Pathway triggered by stimuli treatment, enforced gene expression and gene knockdown.
Principle:
The p53 pathway plays a crucial role in effective tumor suppression because of its central function in cell cycle regulation, DNA repair, cellular senescence, and apoptosis, which can be used for potentially develop new drug therapies against cancer. Upon activation by DNA damage, oncogene activation, or hypoxia, p53 binds to its DNA recognition site on the promoter regions of the target genes and regulate the gene expression.?
Signosis
has developed p53 luciferase reporter Hela stable cell line, in which luciferase activity is specifically associated with the activity of p53. Therefore, the cell line can be used as a reporter system for monitoring the activation of p53 triggered by stimuli treatment, enforced gene expression and gene knockdown.

The cell line was established by transfection?using a?pTA-p53-luciferase reporter vector, which contains 6 repeats of p53 binding sites, a minimal promoter upstream of the firefly luciferase coding region,?along with hygromycin expression vector followed by hygromycin selection.?The hygromycin resistant clones were subsequently screened for doxorubicin-induced luciferase activity.

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Principle behind TF luciferase reporter.
? TF luciferase reporter stable cell line utilizes artificial promoter constructs to drive luciferase expression.? The promoter region can consists of multiple repeats of a cis-element TF binding site, a DNA fragment from the promoter region of a known TF downstream gene, or a DNA fragment containing putative/known TF binding sites.? There are several ways that a TF can be activated, such as through extracellular stimuli or through intracellular signaling pathways.? Once activated, the TF translocates to the nucleus and often interacts with relevant co-factors to drive gene expression.? Once luciferase is expressed, it can generate light in an enzymatic assay and the amount of light measured is positively correlated with the level of TF activation.





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Data:

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Analysis of p53 Reporter Hela Stable Cell Line in response to stimuli.?
The cells were seeded on a 96-well plate for overnight with DMEM including 10% FBS. The cells then were either treated with 500ng/ml Doxorubicin or 2ug/ml Quinacrine respectively in DMEM and 10% FBS for 16 hours.