Nrf2 Transcription Factor Assay Kit (Colorimetric) (ab207223)
Key features and details
- Assay type: Semi-quantitative
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 3 hr 30 min
- Sample type: Nuclear Extracts
- Sensitivity: 600 ng/well
Overview
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Product name
Nrf2 Transcription Factor Assay Kit (Colorimetric)
See all Nrf2 kits -
Detection method
Colorimetric -
Sample type
Nuclear Extracts -
Assay type
Semi-quantitative -
Sensitivity
> 600 ng/well -
Assay time
3h 30m -
Species reactivity
Reacts with: Mouse, Rat, Human -
Product overview
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 µg nuclear extract/well.
Detection range: 0.6 – 10 µg nuclear extract/well.
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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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Platform
Microplate reader
Properties
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Storage instructions
Please refer to protocols. -
Components 1 x 96 tests 5 x 96 tests 10X Antibody Binding Buffer 1 x 2.2ml 5 x 2.2ml 10X Wash Buffer 1 x 22ml 5 x 22ml 96-well Nrf2 assay plate 1 unit 5 units Anti-rabbit HRP-conjugated IgG 1 x 10µl 5 x 10µl Binding Buffer 1 x 10ml 5 x 10ml Developing Solution 1 x 11ml 5 x 11ml Dithiothreitol (DTT) (1 M) 1 x 100µl 5 x 100µl Herring sperm DNA 1 x 100µl 5 x 100µl Lysis Buffer 1 x 10ml 5 x 10ml Mutated oligonucleotide (10 pmol/µL) 1 x 100µl 5 x 100µl Nrf2 antibody 1 x 10µl 1 x 25µl Plate sealer 1 unit 5 units Positive control extract (2.5 µg/µL) 1 x 20µl 1 x 50µl Protease Inhibitor Cocktail 1 x 100µl 5 x 100µl Stop Solution 1 x 11ml 5 x 11ml Wild-type oligonucleotide (10 pmol/µL) 1 x 100µl 5 x 100µl -
Research areas
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Function
Transcription activator that binds to antioxidant response (ARE) elements in the promoter regions of target genes. Important for the coordinated up-regulation of genes in response to oxidative stress. May be involved in the transcriptional activation of genes of the beta-globin cluster by mediating enhancer activity of hypersensitive site 2 of the beta-globin locus control region. -
Tissue specificity
Widely expressed. Highest expression in adult muscle, kidney, lung, liver and in fetal muscle. -
Sequence similarities
Belongs to the bZIP family. CNC subfamily.
Contains 1 bZIP domain. -
Domain
Acidic activation domain in the N-terminus, and DNA binding domain in the C-terminus. -
Post-translational
modificationsPhosphorylation of Ser-40 by PKC in response to oxidative stress dissociates NFE2L2 from its cytoplasmic inhibitor KEAP1, promoting its translocation into the nucleus. -
Cellular localization
Cytoplasm > cytosol. Nucleus. Cytosolic under unstressed conditions, translocates into the nucleus upon induction by electrophilic agents. - Information by UniProt
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Alternative names
- erythroid derived 2
- HEBP1
- like 2
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Database links
- Entrez Gene: 4780 Human
- Entrez Gene: 18024 Mouse
- Entrez Gene: 83619 Rat
- Omim: 600492 Human
- SwissProt: Q16236 Human
- SwissProt: Q60795 Mouse
- SwissProt: O54968 Rat
- Unigene: 1025 Mouse
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Images
Datasheets and documents
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SDS download
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Datasheet download
References (30)
ab207223 has been referenced in 30 publications.
- Kumar B et al. Tumor collection/processing under physioxia uncovers highly relevant signaling networks and drug sensitivity. Sci Adv 8:eabh3375 (2022). PubMed: 35020422
- Xie W et al. Betulinic acid accelerates diabetic wound healing by modulating hyperglycemia-induced oxidative stress, inflammation and glucose intolerance. Burns Trauma 10:tkac007 (2022). PubMed: 35415192
- Meng J & Xu C MicroRNA-495-3p diminishes doxorubicin-induced cardiotoxicity through activating AKT. J Cell Mol Med 26:2076-2088 (2022). PubMed: 35152537
- Asaad GF & Mostafa RE Lactoferrin mitigates ethanol-induced gastric ulcer via modulation of ROS/ICAM-1/Nrf2 signaling pathway in Wistar rats. Iran J Basic Med Sci 25:1522-1527 (2022). PubMed: 36544526
- Kabel AM et al. Effect of dapagliflozin and/or L-arginine on solid tumor model in mice: The interaction between nitric oxide, transforming growth factor-beta 1, autophagy, and apoptosis. Fundam Clin Pharmacol 35:968-978 (2021). PubMed: 33609317