Validation Data Gallery
|Positive WB detected in||4T1 cells, HSC-T6 cells, NIH/3T3 cells|
|Positive IHC detected in||human pancreas cancer tissue, human kidney tissue, human skeletal muscle tissue|
Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
|Western Blot (WB)||WB : 1:1000-1:4000|
|Immunohistochemistry (IHC)||IHC : 1:50-1:500|
|Sample-dependent, check data in validation data gallery|
60027-1-Ig targets KEAP1 in WB, IHC, IF, ELISA applications and shows reactivity with human, rat, mouse samples.
|Tested Reactivity||human, rat, mouse|
|Cited Reactivity||human, mouse, rat, pig|
|Host / Isotype||Mouse / IgA|
|Immunogen||KEAP1 fusion protein Ag0779|
|Full Name||kelch-like ECH-associated protein 1|
|Calculated molecular weight||624 aa, 70 kDa|
|Observed molecular weight||70 kDa|
|GenBank accession number||BC002930|
|Gene ID (NCBI)||9817|
|Purification Method||Thiophilic affinity chromatograph|
|Storage Buffer||PBS with 0.1% sodium azide and 50% glycerol pH 7.3.|
|Storage Conditions||Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage.|
Kelch-like ECH-associated protein 1 (KEAP1) is a negative regulator of nuclear factor erythroid 2-related factor 2 (Nrf2), a transcription factor governing the antioxidant response.
What is the molecular weight of KEAP1 protein? Are there any isoforms of KEAP1?
The molecular weight of KEAP1 protein is 70 kDa. The KEAP1 gene gives rise only to protein isoforms, but mutations of KEAP1 protein have been found in various cancer types.
What is the subcellular localization of KEAP1?
KEAP1 resides in the cytoplasm, where it binds to Nrf2, targeting it for degradation and preventing translocation of Nrf2 to the nucleus.
How does KEAP1 control Nrf2 levels? Is KEAP1 post-translationally modified?
KEAP1 is rich in reactive cysteine residues, whose thiol groups play a role in binding to CUL3 and the polyubiquitination of Nrf2, which leads to degradation of Nrf2 via the proteasome system. During oxidative stress, electrophiles and reactive oxygen species (ROS) modify the KEAP1 thiol groups, reducing the affinity of KEAP1 to CUL3 and the stabilization of Nrf2. Nrf2 then translocates to the nucleus, where it binds to the antioxidant responsive elements (AREs) and induces the expression of antioxidant proteins (PMID: 16354693).
How to measure oxidative stress using KEAP1 and Nrf2 proteins as a readout
Under basal conditions (unstressed cells), a detectable KEAP1 protein level is observed. Oxidative stress modifies KEAP1 protein activity by increasing the Nrf2 protein levels. This can be measured, for example, using western blotting (PMID: 27697860). KEAP1 protein levels are not altered by oxidative stress.
What is the role of the KEAP1-Nrf2 pathway in health and disease?
The KEAP-Nrf2 pathway plays a vital role in redox homeostasis and cryoprotection. Inhibition of KEAP1 activity leads to the activation of Nrf2 and increase the response to oxidative stress and anti-inflammatory effects (PMID: 29717933). The activation of Nrf2 can be beneficial in the case of metabolic diseases, such as diabetes, as well as neurodegenerative diseases such as Parkinson's and Alzheimer's diseases. However, the increased activation of Nrf2 is also known to promote tumor growth and metastasis. Mutations in both KEAP1 and Nrf2 were found in various solid tumor types.
Arch Plast Surg
Expression of nuclear factor erythroid 2 protein in malignant cutaneous tumors.
Pathol Oncol Res
Expression of LC3, LAMP2, KEAP1 and NRF2 in Salivary Adenoid Cystic Carcinoma.
Acta Neuropathol Commun
Activation of the Keap1/Nrf2 stress response pathway in autophagic vacuolar myopathies.
Oxid Med Cell Longev
Neuroprotective Effect of Salvianolic Acid A against Diabetic Peripheral Neuropathy through Modulation of Nrf2.
Int J Nanomedicine
La2O3 Nanoparticles Induce Reproductive Toxicity Mediated by the Nrf-2/ARE Signaling Pathway in Kunming Mice.
Oxid Med Cell Longev
Role of Nrf2 in Lipopolysaccharide-Induced Acute Kidney Injury: Protection by Human Umbilical Cord Blood Mononuclear Cells.
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