• Featured Product
  • KD/KO Validated

KEAP1 Polyclonal antibody

KEAP1 Polyclonal Antibody for IF, IHC, IP, WB, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse and More (6)

Applications

WB, IP, IHC, IF, CoIP, ELISA

Conjugate

Unconjugated

Cat no : 10503-2-AP

Synonyms

Cytosolic inhibitor of Nrf2, INrf2, KEAP1, Kelch like protein 19, KIAA0132, KLHL19



Tested Applications

Positive WB detected inHEK-293 cells, HepG2 cells, Jurkat cells, MCF-7 cells, HeLa cells
Positive IP detected inmouse skeletal muscle tissue
Positive IHC detected inhuman lung cancer tissue, human breast cancer 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
Positive IF detected inU2OS cells, HepG2 cells

Recommended dilution

ApplicationDilution
Western Blot (WB)WB : 1:2000-1:10000
Immunoprecipitation (IP)IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
Immunohistochemistry (IHC)IHC : 1:50-1:500
Immunofluorescence (IF)IF : 1:50-1:500
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, check data in validation data gallery

Product Information

10503-2-AP targets KEAP1 in WB, IP, IHC, IF, CoIP, ELISA applications and shows reactivity with human, mouse samples.

Tested Reactivity human, mouse
Cited Reactivityhuman, mouse, rat, chicken, yellow catfish, monkey, pig, bovine
Host / Isotype Rabbit / IgG
Class Polyclonal
Type Antibody
Immunogen KEAP1 fusion protein Ag0779
Full Name kelch-like ECH-associated protein 1
Calculated molecular weight 624 aa, 70 kDa
Observed molecular weight 55~70 kDa
GenBank accession numberBC002930
Gene symbol KEAP1
Gene ID (NCBI) 9817
RRIDAB_2132625
Conjugate Unconjugated
Form Liquid
Purification Method Antigen affinity purification
Storage Buffer PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
Storage ConditionsStore at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage. 20ul sizes contain 0.1% BSA.

Background Information

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.

Protocols

Product Specific Protocols
WB protocol for KEAP1 antibody 10503-2-APDownload protocol
IHC protocol for KEAP1 antibody 10503-2-APDownload protocol
IF protocol for KEAP1 antibody 10503-2-APDownload protocol
IP protocol for KEAP1 antibody 10503-2-APDownload protocol
Standard Protocols
Click here to view our Standard Protocols

Publications

SpeciesApplicationTitle
humanWB

Nature

KLHL22 activates amino-acid-dependent mTORC1 signalling to promote tumorigenesis and ageing.

Authors - Jie Chen
  • KO Validated
humanWB,CoIP

Cell

Systematic discovery of mutation-directed neo-protein-protein interactions in cancer.

Authors - Xiulei Mo
  • KD Validated
humanWB

Cell

Nrf2 Activation Promotes Lung Cancer Metastasis by Inhibiting the Degradation of Bach1.

Authors - Luca Lignitto
humanWB

Cell

Hypoxia Rescues Frataxin Loss by Restoring Iron Sulfur Cluster Biogenesis.

Authors - Tslil Ast
humanWB, IP

Cancer Cell

iASPP Is an Antioxidative Factor and Drives Cancer Growth and Drug Resistance by Competing with Nrf2 for Keap1 Binding.

Authors - Wenjie Ge
  • KD Validated
human,mouseWB,IF

Nat Cell Biol

The selective autophagy substrate p62 activates the stress responsive transcription factor Nrf2 through inactivation of Keap1.

Authors - Komatsu Masaaki M

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive forproviding their feedback.


FH

Angie (Verified Customer) (07-06-2023)

The antibody was used at dilution of 1:10 000 incubated at room temperature for 1 hour followed by another hour of incubation with secondary antibody donkey-anti-rabbit (Alexa Fluor 800, 1:20 000).

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:10 000
  • Cell Tissue Type: HeLa, MCF7 A375, MM415
KEAP1 Antibody Western Blot validation (1:10 000 dilution) in HeLa, MCF7 A375, MM415 (Cat no:10503-2-AP)
FH

james (Verified Customer) (01-04-2021)

very good quick service

  • Applications: Cell culture
FH

Aurélie (Verified Customer) (09-01-2019)

Lysats from Mouse Cortex were loaded on 8% SDS-PAGE and immunobloted using anti-KEAP1 antibody (1/8000) over-night at 4°C.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1/8000
  • Cell Tissue Type: Mouse Cortex
KEAP1 Antibody Western Blot, validation (1/8000 dilution) in Mouse Cortex (Cat no:10503-2-AP)
FH

Mark (Verified Customer) (02-06-2019)

A doublet band appeared at the correct size and this was the only band that appeared on the blot. We did not perform genetic knockout/expression to confirm the specificity of this band, however it appears correct.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:1000
  • Cell Tissue Type: Human breast cancer cells
KEAP1 Antibody Western Blot, validation (1:1000 dilution) in Human breast cancer cells (Cat no:10503-2-AP)