Product Name Catalog # Price   Qty
AP-1 Luciferase Reporter Hela Stable Cell Line SL-0019-FP $2,000
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Description:

AP-1 Responsive Luciferase Reporter Hela Stable Cell Line is derived from human cervical cancer, and stably express firefly luciferase reporter gene under the control of AP-1 response element.  This cell line is an ideal cellular model for monitoring the activation of JNK, ERK, MAPK Signaling Receptor Signaling Pathway triggered by stimuli treatment, enforced gene expression and gene knockdown.


Principle

AP-1 plays a key role in the control of cell proliferation, differentiation, transformation, survival and apoptosis.  cJun and Fos dimerize to form AP-1 and this protein complex selectively binds to DNA sequences containing the TPA-responsive elements (TREs) to regulate gene expression.  Many stimuli, both physiological and pathological, can regulate AP-1 activity, including TPA, serum, growth factors, cytokines, stress signals, infections, and oncoproteins, TNF and IL-1.  AP-1 activity can be used as readout for JNK, ERK, or MAPK kinase signaling pathway. 

Signosis has developed AP-1 luciferase reporter stable cell line by transducing cells with baculovirus containing both AP-1 luciferase reporter and hygromycin expression cassette.  The hygromycin resistant clones were subsequently screened for PMA-induced luciferase activity.   The cell line can be used as a reporter system for monitoring the activity of AP-1 triggered by stimuli treatment, such as TNFa, IL-1, gene overexpression and gene knockdown.  The cells contain no viral particles and require handling at biosafety level 1 protocol. 

 

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.

 

Data

 Analysis of AP-1 Luciferase Reporter Hela Stable Cell Line.  The cells were seeded on a 96-well plate for overnight with DMEM including 10% FBS. The cells then were treated with or without 10ng/ml PMA in DMEM and 0.1% FBS for 6 hours.  More than 50 fold increase in luciferase activity was detected when compared to untreated cells.

Literature

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