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AB207223

Nrf2 Transcription Factor Assay Kit (Colorimetric)

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(41 Publications)

Nrf2 Transcription Factor Assay Kit (Colorimetric) (ab207223) is a high throughput assay to quantify Nrf2 activation in nuclear extracts.

View Alternative Names

NRF2, NFE2L2, Nuclear factor erythroid 2-related factor 2, NF-E2-related factor 2, NFE2-related factor 2, Nrf-2

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Functional Studies - Nrf2 Transcription Factor Assay Kit (Colorimetric) (AB207223)
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Supplier Data

Functional Studies - Nrf2 Transcription Factor Assay Kit (Colorimetric) (AB207223)

Nuclear extracts from untreated HepG2 cells (Light gray) and HepG2 cells treated with D,L Sulforaphane (Black) were assayed from 0.625 to 10 µg/well for Nrf2 activation using ab207223. Different amounts of nuclear extracts from untreated HepG2 cells (light grey) and HepG2 cells treated with D,L-Sulforaphane (Black) were tested for Nrf2 activation. These results are provided for demonstration only.

Key facts

Detection method

Colorimetric

Sample types

Nuclear Extracts

Reacts with

Mouse, Rat, Human

Assay type

Semi-quantitative

Sensitivity

> 600 ng/well

Assay time

3h 30m

Assay Platform

Microplate reader

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Product details

Nrf2 Transcription Factor Assay Kit (Colorimetric) (ab207223) is a high throughput assay to quantify Nrf2 activation in nuclear extracts. This assay combines a quick ELISA format with a sensitive and specific non-radioactive assay for transcription factor activation.

A specific double stranded DNA sequence containing the Nrf2 consensus binding site (5'-GTCACAGTGACTCAGCAGAATCTG-3') has been immobilized onto a 96-well plate. Active Nrf2 present in the nuclear extract specifically binds to the oligonucleotide. Nrf2 is detected by a primary antibody that recognizes an epitope of Nrf2 accessible only when the protein is activated and bound to its target DNA. An HRP-conjugated secondary antibody provides sensitive colorimetric readout at OD 450 nm. This product detects only human, mouse and rat Nrf2.

Key performance and benefits:

Assay time: 3.5 hours (cell extracts preparation not included).

Detection limit: < 0.6 ug nuclear extract/well.

Detection range: 0.6 - 10 ug nuclear extract/well.

Other Notes
Nrf2 (NF-E2 related factor, NFE2L2, from nuclear factor erythroid-derived 2-like 2) is a basic leucine zipper (bZIP) transcription factor. Nrf2 binds to the antioxidant response element (ARE) and positively regulates the expression of detoxifying enzyme genes (such as NAD(P)H:quinone oxidoreductase1, NQO1) in response to antioxidants and xenobiotics. Higher levels of NQO1 gene expression has been shown in liver, lung, colon, and breast tumors.

A cytosolic inhibitor of Nrf2, Keap1/INrf2, retains Nrf2 in the cytoplasm under normal conditions where the interaction of Nrf2 with INrf2 targets Nrf2 for ubiquitination and proteasomal degradation. However, after oxidative stress, Nrf2 is released from INrf2, translocates to the nucleus, and results in the activation of ARE-mediated gene expression. Nrf2 is also synthesized de novo after exposure to stress. In addition, Nrf2 controls its own degradation by regulating expression and induction of INrf2.

It has been shown that nuclear export and degradation pathways are activated by around two hours after treatment with tert-butylhydroquinone (t-BHQ).

Nrf2 activation and degradation are important sensing mechanisms in the cellular response for oxidative and electrophilic stressors.

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What's included?

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Properties and storage information

Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
Multi
Appropriate long-term storage conditions
Multi
Storage information
Please refer to protocols
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Nrf2 also known as nuclear factor erythroid 2-related factor 2 is a transcription factor with a molecular weight of approximately 66 kDa. It plays a mechanical role in regulating the expression of antioxidant proteins that protect against oxidative damage triggered by injury and inflammation. Nrf2 is ubiquitously expressed in various tissues including the liver lungs and skin. Nrf2 activation occurs when it translocates from the cytoplasm to the nucleus to bind the antioxidant response element (ARE) in the DNA starting transcription of target genes.
Biological function summary

Nrf2 acts as an important regulator of the cellular antioxidant response. It works in conjunction with its partner protein Keap1 forming a complex that controls its stability and degradation. Under normal conditions Keap1 keeps Nrf2 in the cytoplasm where it is targeted for ubiquitination and degradation. Once activated by oxidative stress or electrophiles Nrf2 dissociates from Keap1 thereby avoiding degradation and relocates to the nucleus to activate the transcription of ARE-dependent genes. This activity boosts the cellular response to oxidative stress by inducing genes involved in detoxification and cellular defense.

Pathways

Nrf2 plays a significant role in the oxidative stress response and detoxification pathways. Nrf2 activation is linked closely to the PI3K/Akt signaling pathway which influences cell survival growth and metabolism. This pathway also interacts with other important proteins like GSK-3β which can modulate Nrf2 activity and stability. Through these pathways Nrf2 orchestrates a defense mechanism against reactive oxygen species (ROS) by boosting the expression of antioxidant enzymes and detoxifying proteins.

Nrf2 has been associated with conditions like cancer and neurodegenerative diseases. In cancer aberrant Nrf2 activation may lead to enhanced tumor survival by increasing expression of cytoprotective genes which makes cancer cells resistant to chemotherapy. Nrf2 also interacts with proteins such as p53 which play roles in tumor suppression and cellular stress responses. In neurodegenerative disorders reduced Nrf2 activity can contribute to oxidative stress leading to neuron damage and disease progression with proteins like amyloid-beta also being linked with oxidative processes affected by Nrf2 functionality.
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Product protocols

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Target data

The protein expressed by the NFE2L2 gene functions as a transcription factor that plays a key role in responding to oxidative stress by binding to antioxidant response elements (ARE) in the promoter regions of various cytoprotective genes, such as phase 2 detoxifying enzymes, to promote their expression and neutralize reactive electrophiles. Under normal conditions, it is ubiquitinated and degraded in the cytoplasm by the BCR(KEAP1) complex. However, oxidative stress leads to the inhibition of the BCR(KEAP1) complex, allowing NFE2L2/NRF2 to accumulate in the nucleus, form heterodimers with small Maf proteins, and bind to ARE elements. Additionally, the NFE2L2/NRF2 pathway is activated by selective autophagy, where KEAP1 interacts with SQSTM1/p62, inactivating the BCR(KEAP1) complex and leading to nuclear NFE2L2/NRF2 accumulation and cytoprotective gene expression. It may also be involved in activating genes of the beta-globin cluster by facilitating enhancer activity of hypersensitive site 2 of the beta-globin locus control region. This supplementary information is collated from multiple sources and compiled automatically.
See full target information NFE2L2
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Publications (41)

Recent publications for all applications. Explore the full list and refine your search

Brain sciences 13: PubMed38137165

2023

Differential Effects of Paraquat, Rotenone, and MPTP on Cellular Bioenergetics of Undifferentiated and Differentiated Human Neuroblastoma Cells.

Applications

Unspecified application

Species

Unspecified reactive species

Ekramy Elmorsy,Ayat Al-Ghafari,Huda Al Doghaither,Sara Hashish,Mohamed Salama,Anusha W Mudyanselage,Lipta James,Wayne G Carter

World journal of stem cells 15:1017-1034 PubMed38058959

2023

Dissecting molecular mechanisms underlying ferroptosis in human umbilical cord mesenchymal stem cells: Role of cystathionine γ-lyase/hydrogen sulfide pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Bin Hu,Xiang-Xi Zhang,Tao Zhang,Wan-Cheng Yu

BMC pharmacology & toxicology 24:63 PubMed37986186

2023

Bisdemethoxycurcumin alleviates LPS-induced acute lung injury via activating AMPKα pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Huifang Li,Qi Zou,Xueming Wang

Cell reports. Medicine 4:101157 PubMed37586364

2023

Boosting NAD preferentially blunts Th17 inflammation via arginine biosynthesis and redox control in healthy and psoriasis subjects.

Applications

Unspecified application

Species

Unspecified reactive species

Kim Han,Komudi Singh,Allison M Meadows,Rahul Sharma,Shahin Hassanzadeh,Jing Wu,Haley Goss-Holmes,Rebecca D Huffstutler,Heather L Teague,Nehal N Mehta,Julian L Griffin,Rong Tian,Javier Traba,Michael N Sack

Cell death & disease 14:372 PubMed37355685

2023

TRIB1 regulates liver regeneration by antagonizing the NRF2-mediated antioxidant response.

Applications

Unspecified application

Species

Unspecified reactive species

Xinyue Sun,Shuai Wang,Xiulian Miao,Sheng Zeng,Yan Guo,Anqi Zhou,Ying Chen,Yifei Chen,Fangqiao Lv,Zhiwen Fan,Yutong Wang,Yong Xu,Zilong Li

Antioxidants (Basel, Switzerland) 12: PubMed36979014

2023

Photobiomodulation Controls Keratinocytes Inflammatory Response through Nrf2 and Reduces Langerhans Cells Activation.

Applications

Unspecified application

Species

Unspecified reactive species

Sara Salman,Cyprien Guermonprez,Laurent Peno-Mazzarino,Elian Lati,Audrey Rousseaud,Lieve Declercq,Saadia Kerdine-Römer

ACS omega 8:7666-7683 PubMed36872984

2023

Terminators or Guardians? Design, Synthesis, and Cytotoxicity Profiling of Chalcone-Sulfonamide Hybrids.

Applications

Unspecified application

Species

Unspecified reactive species

Shaimaa M Aboukhatwa,Peter A Sidhom,Andrea Angeli,Claudiu T Supuran,Haytham O Tawfik

International journal of molecular sciences 24: PubMed36769034

2023

Moderating Gut Microbiome/Mitochondrial Axis in Oxazolone Induced Ulcerative Colitis: The Evolving Role of β-Glucan and/or, Aldose Reductase Inhibitor, Fidarestat.

Applications

Unspecified application

Species

Unspecified reactive species

Omnia Safwat El-Deeb,Rasha Osama Elesawy,Amira K Eltokhy,Hanan Alsaeid Al-Shenawy,Heba Bassiony Ghanem,Fatma H Rizk,Ramez Ae Barhoma,Rania H Shalaby,Amal M Abdelsattar,Shaimaa S Mashal,Kareman Ahmed Eshra,Radwa Mahmoud El-Sharaby,Dina Adam Ali,Rowida Raafat Ibrahim

Iranian journal of basic medical sciences 25:1522-1527 PubMed36544526

2022

Lactoferrin mitigates ethanol-induced gastric ulcer via modulation of ROS/ICAM-1/Nrf2 signaling pathway in Wistar rats.

Applications

Unspecified application

Species

Unspecified reactive species

Gihan F Asaad,Rasha E Mostafa

British journal of pharmacology 180:308-329 PubMed36166825

2022

Icariside II preconditioning evokes robust neuroprotection against ischaemic stroke, by targeting Nrf2 and the OXPHOS/NF-κB/ferroptosis pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Jianmei Gao,Congjian Ma,Dianya Xia,Nana Chen,Jianyong Zhang,Fan Xu,Fei Li,Yuqi He,Qihai Gong
View all publications
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