商品编号


SL-0006-NP



品牌


Signosis



公司


Signosis



公司分类


Cell-Based Assays




1 Ea


商品信息

Description:
NFkB Responsive Luciferase Reporter NIH/3T3 Stable Cell Line is derived from mouse fibroblast,and stably express firefly luciferase reporter gene under the control of the NFkB??Response element.This cell line is an ideal cellular model for monitoring the activation of NFkB Receptor Signaling Pathway triggered by stimuli treatment, enforced gene expression and gene knockdown.
Principle:
NF
k
B plays an important role in controlling many
BIOLOG
ical processes including immune and inflammatory responses, developmental processes, cellular growth, and apoptosis. In response to the various stimuli, such as stress, cytokines, free r
ADI
cals, ultraviolet irr
ADI
ation, and bacterial or viral antigens, NF
k
B is activated and translocates from cytoplasm to nucleus, where NF
k
B binds to its response element on the promoter region and regulates a wide spectrum of gene expression. Dysfunction of NF
k
B activity is associated with cancer, inflammatory and autoimmune disease, and viral infection. Monitoring the NF
k
B activity is essential to unveil the mechanism of these diseases and conduct drug discovery.
Signosis
has established a NF
k
B luciferase reporter stable cell line that has been stably transfected with pTA-NF
k
B-luciferase reporter vector and?can be used as a reporter system for monitoring the activation of NF
k
B triggered by stimuli treatment?such as TNFα, IL-1α and IL-1β, enforced gene expression and gene knockdown.?

The cell line was established by transfection?using a?pTA-NFkB-luciferase reporter vector, which contains 4 repeats of NFkB 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 TNFa-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 SL-0006 NFκB reporter activity in response to various treatments.?
The cells were seeded on a 96-well plate for overnight with DMEM including 10% FBS. The cells then were treated with the following chemicals in DMEM and 0.1% FBS for 16 hours: 10ng/mL IL-1a, , 10ng/ml PMA, 10ng/ml oncostatin M (OSM), 100ng/ml IFN-gamma, and 2ng/ml TNFα.? NIH/3T3-NFkB Luciferase Reporter Cell Line exhibits response to TNFα and IL-1a when compared to untreated cells.