Product Name Catalog # Price (NP)**   Qty
NFAT Luciferase Reporter Jurkat T Stable Cell Line SL-0032-NP $950
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  • ** Non Profit (NP) price is for academic, non profit organizations and institutes
Description:

NFAT  Responsive Luciferase Reporter Jurkat T Stable Cell Line is derived from human T lymphocyte ,and stably express firefly luciferase reporter gene under the control of NFAT response element.  This cell line is an ideal cellular model for monitoring the activation of Calcium Signaling Pathway triggered by stimuli treatment, enforced gene expression and gene knockdown.


Principle

T cell receptor signaling plays an important role in T cell activation, proliferation and differentiation leading to the initiation series tyrosine phosphorylation events that trigger multiple signaling pathways, one of which is the activation of Ca2+/protein kinase C pathways. The increased intracellular calcium levels and the activation of PKCs leads to the activation the nuclear factor of activator T cells (NFAT) transcription factor resulting in downstream gene expression. Jurkat leukaemic T-cell line is widely used  cell line for  studying TCR/NFAT signaling.  Signosis’ Jurkat NFAT Reporter Stable Cell Line is established by transfection using a pTA-NFAT-luciferase reporter vector, which contains NFAT 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 luciferase activityinduced by PMA+ionomycin.

 

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-0032

 

 

 

 

 

 

 

 

 

 

Analysis of NFAT Luciferase Reporter Jurkat T Stable Cell Line. The cells were seeded on  a 96-well plate in media containing 10ng/ml PMA, 1uM ionomycin, and 0.1% FBS for 16 hours.  More than 200 fold increase in luciferase activity was detected when compared to untreated cells.

Literature

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