Product Name Catalog # Price   Qty
NFκB Luciferase Reporter HeLa Stable Cell Line SL-0001-FP $2,000
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Description:

NFkB  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 NFkB response element. This cell line is an ideal cellular model for monitoring the activation of NFkB Pathway triggered by stimuli treatment, enforced gene expression and gene knockdown.


Principle

NFkB plays an important role in controlling many biological processes including immune and inflammatory responses, developmental processes, cellular growth, and apoptosis. In response to the various stimuli, such as stress, cytokines, free radicals, ultraviolet irradiation, and bacterial or viral antigens, NFkB is activated and translocates from cytoplasm to nucleus, where NFkB binds to its response element on the promoter region and regulates a wide spectrum of gene expression. Dysfunction of NFkB activity is associated with cancer, inflammatory and autoimmune disease, and viral infection. Monitoring the NFkB activity is essential to unveil the mechanism of these diseases and conduct drug discovery. Signosis has established an NFkB luciferase reporter stable cell line that has been stably transfected with pTA-NFkB-luciferase reporter vector, which contains 4 repeats of NFkB binding sites, a minimal promoter upstream of the firefly luciferase coding region. Therefore, the cell line can be used as a reporter system for monitoring the activation of NFkB triggered by stimuli treatment, enforced gene expression and gene knockdown.  The stable cell line can be used for studying NFkB signaling pathways activated by different cytokines, such as TNFa, and many other stimuli. 

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 (or IL-1) 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

Analysis of TNFa/NFkB Pathway Reporter Cell Line in response to TNFa treatment.    The cells were seeded on  a 96-well plate for overnight with DMEM including 10% FBS. The cell then were treated with 20ng/ml TNFa in DMEM + 0.1% FBS for 6 hours. 

Analysis of TNFa/NFkB Pathway Reporter Cell Line in response to TNFa treatment.   
The cells were seeded on  a 96-well plate for overnight with DMEM including 10% FBS. The cell then were treated with 20ng/ml TNFa in DMEM + 0.1% FBS for 6 hours.

 

Literature

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Citations

 

NRF2/ARE Luciferase Reporter MCF7 Stable Cell Line  SL-0010 
 
Withaferin A induces Nrf2-dependent protection against liver injury: role of Keap1-independent mechanisms. DL Palliyaguru, DV Chartoumpekis, N Wakabayashi. Free Radical Biology and Medicine 2016. http://dx.doi.org/10.1016/j.freeradbiomed.2016.10.003
 
Luciferase Stable Expressing Hela Cell Line SL-0102
 
Redox-responsive, reversibly-crosslinked thiolated cationic helical polypeptides for efficient siRNA encapsulation and delivery.N Zheng, Z Song, Y Liu, R Zhang, R Zhang, C Yao. Journal of Controlled Release.Volume 205, 10 May 2015, Pages 231–239
 
Targeted delivery of cisplatin to tumor xenografts via the nanoparticle component of nano-diamino-tetrac. Thangirala Sudha, Dhruba J Bharali, Murat Yalcin, Noureldien HE Darwish, Melis Debreli Coskun, Kelly A Keating, Hung-Yun Lin, Paul J Davis, Shaker A Mousa. February 2017 ,Vol. 12, No. 3, Pages 195-205 , DOI 10.2217/nnm-2016-0315.
 
NFkB Luciferase Reporter Stable Cell Lines  SL-0001
 
Thymoquinone Modulates Blood Coagulation in Vitro via Its Effects on Inflammatory and Coagulation Pathways. V Muralidharan-Chari, J Kim, A Abuawad, M Naeem.  Int. J. Mol. Sci. 2016, 17(4), 474; doi:10.3390/ijms17040474
 
Drug repurposing screen identifies lestaurtinib amplifies the ability of the poly (ADP-ribose) polymerase 1 inhibitor AG14361 to kill breast cancer associated gene-1 …G Vazquez-Ortiz, C Chisholm, X Xu, TJ Lahusen, C Li… - Breast Cancer Research  2014,  Jun 24;16(3):R67. doi: 10.1186/bcr3682.
 
NFkB Luciferase Reporter A549 Stable Cell Lines  SL-0014
 
Alcohol promotes migration and invasion of triple-negative breast cancer cells through activation of p38 MAPK and JNK†M Zhao, EW Howard, AB Parris, Z Guo, Q Zhao, X Yang.  Molecular Carcinogenesis, 2016, DOI: 10.1002/mc.22538
 
HEK293 TCF/LEF Luciferase Reporter Stable Cell Line SL-0015
 
Active β-catenin is regulated by the PTEN/PI3 kinase pathway: a role for protein phosphatase PP2A. Amit Persad,1,* Geetha Venkateswaran,1,* Li Hao,1 Maria E. Garcia,1 Jenny Yoon,1 Jaskiran Sidhu,1 and Sujata Persad1. Genes Cancer. 2016 Nov; 7(11-12): 368–382., doi:  10.18632/genesandcancer.128
 
Diallyl trisulfide inhibits proliferation, invasion and angiogenesis of glioma cells by inactivating Wnt/β-catenin signaling. Qingxia TaoCuiying WuRuxiang XuLijun NiuJiazhen QinNing LiuPeng ZhangEmail authorChong Wang. Cell and Tissue Research, 2017, doi:10.1007/s00441-017-2678-9
 
BCN057 induces intestinal stem cell repair and mitigates radiation-induced intestinal injury. Payel Bhanja, Andrew Norris, Pooja Gupta-Saraf, Andrew Hoover and Subhrajit Saha. Stem Cell Research & Therapy, vol. 9, no. 1, Feb. 2018, doi:10.1186/s13287-017-0763-3.
 
(Nrf2/ARE) luciferase reporter cell line HEK293
 
Direct Antioxidant Properties of Methotrexate: Inhibition of Malondialdehyde-Acetaldehyde-Protein Adduct Formation and Superoxide Scavenging. Matthew C. Zimmermana, Dahn L. Clemensb, c, Michael J. Duryeeb, Cleofes Sarmientoa, Andrew Chioub, Carlos D. Hunterb, Jun Tiana, Lynell W. Klassenb, c, James R. O’Dellb, c, Geoffrey M. Thieleb, c, Redox Biology,2017.07.018, doi: 10.1016/j.redox.2017.07.018
 
Active β-catenin is regulated by the PTEN/PI3 kinase pathway: a role for protein phosphatase PP2A. Amit Persad1,*, Geetha Venkateswaran1,*, Li Hao1, Maria E. Garcia1, Jenny Yoon1, Jaskiran Sidhu1 and Sujata Persad1.Genes & Cancer. January 30, 2017 . https://doi.org/10.18632/genesandcancer.128