Anticorps Polyclonal de lapin anti-KEAP1

• Phare
• Validé par KD/KO

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

Hôte / Isotype

Lapin / IgG

Réactivité testée

Humain, souris et plus (5)

Applications

WB, IP, IHC, IF, CoIP, ELISA

Conjugaison

Non conjugué

Publications

551

Avis (4)

N° de cat : 10503-2-AP

Synonymes

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

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Galerie de données de validation

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  WB analysis using 10503-2-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 10503-2-AP (KEAP1 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  WB analysis of HepG2 cells using 10503-2-AP

  WB result of KEAP1 antibody (10503-2-AP, 1:2000) with si-control and si-KEAP1 transfected HEK293 cell.

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  WB analysis of HEK-293 using 10503-2-AP

  WB result of KEAP1 antibody (10503-2-AP; 1:3000; incubated at room temperature for 1.5 hours) with sh-Control and sh-KEAP1 transfected HEK-293 cells.

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  WB analysis using 10503-2-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 10503-2-AP (KEAP1 antibody) at dilution of 1:6000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 10503-2-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 10503-2-AP (KEAP1 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 10503-2-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 10503-2-AP (KEAP1 antibody) at dilution of 1:8000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 10503-2-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 10503-2-AP (KEAP1 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 10503-2-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 10503-2-AP (KEAP1 antibody) at dilution of 1:15000 incubated at room temperature for 1.5 hours.

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  WB analysis of HEK-293 using 10503-2-AP

  WB result of KEAP1 antibody (10503-2-AP; 1:20000; incubated at room temperature for 1.5 hours) with sh-Control and sh-KEAP1 transfected HEK-293 cells.

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  WB analysis of HEK-293 using 10503-2-AP

  HEK-293 and HepG2 cells were subjected to SDS PAGE followed by western blot with 10503-2-AP (KEAP1 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of Jurkat using 10503-2-AP

  Jurkat cells and tissues were subjected to SDS PAGE followed by western blot with 10503-2-AP (KEAP1 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  IP experiment of mouse skeletal muscle using 10503-2-AP

  IP Result of anti-KEAP1 (IP:10503-2-AP, 5ug; Detection:10503-2-AP 1:600) with mouse skeletal muscle tissue lysate 8000ug.

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  IHC staining of human lung cancer using 10503-2-AP

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 10503-2-AP (KEAP1 Antibody) at dilution of 1:200 (under 10x lens).

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  IHC staining of human lung cancer using 10503-2-AP

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 10503-2-AP (KEAP1 Antibody) at dilution of 1:200 (under 40x lens).

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  IHC staining of human breast cancer using 10503-2-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer using 10503-2-AP (KEAP1 antibody) at dilution of 1:100 (under 25x lens).

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  IHC staining of human skeletal muscle using 10503-2-AP

  Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue slide using 10503-2-AP (KEAP1 Antibody) at dilution of 1:50.

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  IHC staining of human skeletal muscle using 10503-2-AP

  Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue slide using 10503-2-AP (KEAP1 Antibody) at dilution of 1:50.

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  IF Staining of HepG2 using 10503-2-AP

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using KEAP1 antibody (10503-2-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

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  IF Staining of U2OS using 10503-2-AP

  Immunofluorescent analysis of (4% PFA) fixed U2OS cells using KEAP1 antibody (10503-2-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

• 

  IF Staining of HepG2 using 10503-2-AP

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using KEAP1 antibody (10503-2-AP) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-Phallodin (red).

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Applications testées

• Résultats positifs en WB: cellules HEK-293, cellules HeLa, cellules HepG2, cellules Jurkat, cellules MCF-7
• Résultats positifs en IP: tissu de muscle squelettique de souris
• Résultats positifs en IHC: tissu de cancer du poumon humain, tissu de cancer du sein humain, tissu de muscle squelettique humain; il est suggéré de démasquer l'antigène avec un tampon de TE buffer pH 9.0; (*) À défaut, 'le démasquage de l'antigène peut être 'effectué avec un tampon citrate pH 6,0.
• Résultats positifs en IF: cellules U2OS, cellules HepG2

Dilution recommandée

• Western Blot (WB): WB : 1:2000-1:10000
• Immunoprécipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochimie (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

Applications publiées

• KD/KO: See 31 publications below
• WB: See 513 publications below
• IHC: See 39 publications below
• IF: See 55 publications below
• IP: See 25 publications below
• CoIP: See 14 publications below

Informations sur le produit

10503-2-AP cible KEAP1 dans les applications de WB, IP, IHC, IF, CoIP, ELISA et montre une réactivité avec des échantillons Humain, souris

• Réactivité: Humain, souris
• Réactivité citée: rat, bovin, Humain, porc, poulet, singe, souris
• Hôte / Isotype: Lapin / IgG
• Clonalité: Polyclonal
• Type: Anticorps
• Immunogène: KEAP1 Protéine recombinante Ag0779
• Nom complet: kelch-like ECH-associated protein 1
• Masse moléculaire calculée: 624 aa, 70 kDa
• Poids moléculaire observé: 55~70 kDa
• Numéro d’acquisition GenBank: BC002930
• Symbole du gène: KEAP1
• Identification du gène (NCBI): 9817
• Conjugaison: Non conjugué
• Forme: Liquide
• Méthode de purification: Purification par affinité contre l'antigène
• Tampon de stockage: PBS avec azoture de sodium à 0,02 % et glycérol à 50 % pH 7,3
• Conditions de stockage: Stocker à -20°C. Stable pendant un an après l'expédition. L'aliquotage n'est pas nécessaire pour le stockage à -20^oC Les 20ul contiennent 0,1% de BSA.

Informations générales

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.

Publications

Species	Application	Title
human	WB	Nature; KLHL22 activates amino-acid-dependent mTORC1 signalling to promote tumorigenesis and ageing.; Authors - Jie Chen ; KO Validated
human	WB,CoIP	Cell; Systematic discovery of mutation-directed neo-protein-protein interactions in cancer.; Authors - Xiulei Mo ; KD Validated
human	WB	Cell; Nrf2 Activation Promotes Lung Cancer Metastasis by Inhibiting the Degradation of Bach1.; Authors - Luca Lignitto
human	WB	Cell; Hypoxia Rescues Frataxin Loss by Restoring Iron Sulfur Cluster Biogenesis.; Authors - Tslil Ast
human	WB, 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,mouse	WB,IF	Nat Cell Biol; The selective autophagy substrate p62 activates the stress responsive transcription factor Nrf2 through inactivation of Keap1.; Authors - Komatsu Masaaki M

Avis

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

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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)

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