Anticorps Recombinant de lapin anti-KEAP1

• Phare
• Validé par KD/KO

KEAP1 Recombinant Antibody for IF, IHC, WB, ELISA

Hôte / Isotype

Lapin / IgG

Réactivité testée

Humain, rat, souris

Applications

WB, IHC, IF, ELISA

Conjugaison

Non conjugué

Publications

1

CloneNo.

5O18

N° de cat : 80744-1-RR

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 of HEK-293 using 80744-1-RR

  WB result of KEAP1 antibody (80744-1-RR; 1:40000; 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 80744-1-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 80744-1-RR (KEAP1 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  IHC staining of human lung cancer using 80744-1-RR

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 80744-1-RR (KEAP1 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human lung cancer using 80744-1-RR

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 80744-1-RR (KEAP1 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HepG2 using 80744-1-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed HepG2 cells using KEAP1 antibody (80744-1-RR, Clone: 5O18 ) at dilution of 1:1000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

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  IF Staining of HeLa using 80744-1-RR

  Immunofluorescent analysis of (-20°C Methanol) fixed HeLa cells using KEAP1 antibody (80744-1-RR, Clone: 5O18 ) at dilution of 1:1000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

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Applications testées

• Résultats positifs en WB: cellules Jurkat, cellules 4T1, cellules HEK-293, cellules HeLa, cellules HSC-T6, cellules K-562, cellules MCF-7, cellules NIH/3T3, cellules RAW 264.7
• Résultats positifs en IHC: tissu de cancer du poumon 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 HepG2, cellules HeLa

Dilution recommandée

• Western Blot (WB): WB : 1:2000-1:10000
• Immunohistochimie (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF): IF : 1:500-1:2000

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

• WB: See 1 publications below

Informations sur le produit

80744-1-RR cible KEAP1 dans les applications de WB, IHC, IF, ELISA et montre une réactivité avec des échantillons Humain, rat, souris

• Réactivité: Humain, rat, souris
• Hôte / Isotype: Lapin / IgG
• Clonalité: Recombinant
• 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 protéine A
• 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
	WB	Antioxidants (Basel); Effects of Different Dietary Zinc (Zn) Sources on Growth Performance, Zn Metabolism, and Intestinal Health of Grass Carp; Authors - Yi-Chuang Xu