Product Name Catalog # Price   Qty
HIF Luciferase Reporter Neuro2a Stable Cell Line SL-0027-FP $2,000
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Description:

HIF responsive Luciferase Reporter Neuro2a  Stable Cell Line is derived from mouse neuroblastoma, and stably express firefly luciferase reporter gene under the control of HIF response element. This cell line is an ideal cellular model for monitoring the activation of Hypoxia receptor Signaling Pathway triggered by stimuli treatment, enforced gene expression and gene knockdown.


Principle

Hypoxia-Inducible Factor (HIF)-1 is a dimeric protein complex that plays a central role in the response to low oxygen concentrations, or hypoxia, and is a crucial physiological regulator of homeostasis, vascularization, and anaerobic metabolism. HIF is a transcription factor regulating gene expression by binding to their DNA recognition site on the target genes. Furthermore, HIF-1 has been widely studied because of its perceived therapeutic potential.  HIF-1 allows survival and proliferation of cancerous cells due to its angiogenic properties, and the inhibition potentially could prevent the spread of cancer. With a growing understanding of the HIF-1 pathway, it has become an attractive goal to analyze the inhibition and stimulation of HIF transcriptional activity via small molecules. 

Signosis has developed HIF Luciferase Reporter Stable Cell Line by transfection of HIF luciferase reporter vector along with hygromycin expression vector followed by hygromycin selection. The hygromycin resistant clones were subsequently screened for CoCl2 or DMOG-induced luciferase activity.

 

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

SL-0027

 

 

 

 

 

 

 

 

 

Analysis of SL-0027 HIF reporter activity in response to various treatments. The Neuro2a cells were seeded on a 96-well plate for overnight with DMEM including 10% FBS. The cells then were treated with the following chemicals for 16 hours: 100uM DMOG, 100uM CoCl2, 20ng/ml TNFα, and 10ng/ml PMA. Neuro2a-HIF Luciferase Reporter Cell Line exhibits response to DMOG and CoCl2 when compared to untreated cells.