KEAP1 Antibody

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ProSci
Product Code: PSI-7045
Product Group: Primary Antibodies
Supplier: ProSci
CodeSizePrice
PSI-7045-0.02mg0.02mg£150.00
Quantity:
PSI-7045-0.1mg0.1mg£449.00
Quantity:
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Overview

Host Type: Rabbit
Antibody Isotype: IgG
Antibody Clonality: Polyclonal
Regulatory Status: RUO
Applications:
  • Enzyme-Linked Immunosorbent Assay (ELISA)
  • Immunofluorescence (IF)
  • Western Blot (WB)

Images

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<strong>Figure 1 Western Blot Validation of KEAP1 in Human Lung </strong><br> Loading: 15 μg of lysates per lane. Antibodies: KEAP1 7045 (A: 1 μg/mL; B: 2 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: KEAP1 7045 (2 μg/mL), KEAP1 22-002 (2 μg/mL, and GAPHDH (0.02 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 3 KO Validation in HeLa cells</strong><br> Loading: 10 μg of HeLa WT cell lysates or KEAP1 KO cell lysates. Antibodies: KEAP1 7045 (2 μg/mL) and beta-actin 3779 (1 μg/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 4 Immunofluorescence Validation of KEAP1 in Human Lung</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed human lung tissue labeling KEAP1 with 7045 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
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<strong>Figure 1 Western Blot Validation of KEAP1 in Human Lung </strong><br> Loading: 15 μg of lysates per lane. Antibodies: KEAP1 7045 (A: 1 μg/mL; B: 2 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: KEAP1 7045 (2 μg/mL), KEAP1 22-002 (2 μg/mL, and GAPHDH (0.02 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 KO Validation in HeLa cells</strong><br> Loading: 10 μg of HeLa WT cell lysates or KEAP1 KO cell lysates. Antibodies: KEAP1 7045 (2 μg/mL) and beta-actin 3779 (1 μg/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 Immunofluorescence Validation of KEAP1 in Human Lung</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed human lung tissue labeling KEAP1 with 7045 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).

Documents

Further Information

Additional Names:
KEAP1 Antibody: INrf2, KLHL19, INRF2, KIAA0132, Kelch-like ECH-associated protein 1, Cytosolic inhibitor of Nrf2, INrf2
Application Note:
WB: 1-2 μg/mL; IF: 20 μg/mL.

Antibody validated: Western Blot in human, mouse and rat samples; Immunofluorescence in human samples. All other applications and species not yet tested.
Background:
KEAP1 Antibody: KEAP1 (kelch-like ECH-associated protein 1) is a stress sensing adaptor for the Cullin3 (Cul3)-dependent E3 ubiquitin ligase complex that negatively regulates NRF2 (NF-E2-related factor 2) and plays a role in the oxidative stress response. It targets NFE2L2/NRF2 for ubiquitination and degradation by the proteasome. KEAP1 contains an amino terminal BTB/POZ domain and a carboxyl terminal KELCH domain which are required for interaction with NRF2, and in binding Cul3-E3 ubiquitin ligase. Altered expression of NRF2 is associated with chronic obstructive pulmonary disease (COPD). KEAP1 also targets the down regulation of NF-κB activity by targeting IKKβ degradation. Mutation of the KEAP1 gene is found in lung cancer.
Background References:
  • Zhang et al. Mol. Cell. Biol. 2004;24:10941-53.
  • Kobayashi et al. Mol. Cell. Biol. 2004; 24:7130-9.
  • Jiang et al. Int. J. Mol. Med. 2012; 29:946-56.
  • Devling et al. Proc. Natl. Acad. Sci. USA 2005; 102: 7280-5A.
Buffer:
KEAP1 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Conjugate:
Unconjugated
DISCLAIMER:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.
Homology:
Predicted species reactivity based on immunogen sequence: Pig: (100%)
Immunogen:
Anti-KEAP1 antibody (7045) was raised against a 19 amino acid synthetic peptide near the carboxy terminus of human KEAP1.

The immunogen is located within the last 50 amino acids of KEAP1.
ISOFORMS:
Human KEAP1 has 1 isoform, (624aa, 69.7kD). Mouse KEAP1 has 1 isoform (624aa, 69.6kD). Rat KEAP1 has only one isoform (624aa, 69.4 kD). This antibody detects KEAP1 of human, mouse and rat.
NCBI Gene ID #:
9817
NCBI Official Name:
kelch-like ECH-associated protein 1
NCBI Official Symbol:
KEAP1
NCBI Organism:
homo sapiens
Physical State:
Liquid
PREDICTED MOLECULAR WEIGHT:
Predicted: 70kD

Observed: 65-70kD
Protein Accession #:
NP_987096
Protein GI Number:
45269145
Purification:
KEAP1 Antibody is affinity chromatography purified via peptide column.
Research Area:
Signal Transduction
SPECIFICITY:
At least two isoforms of KEAP1 are known to exist.
Swissprot #:
Q14145
User NOte:
Optimal dilutions for each application to be determined by the researcher.
VALIDATION:

Independent Antibody Validation in Cell lines (Figure 2) shows similar KEAP1 expression profile in human, mouse (3T3/NIH) and rat (YB2/0) cell lines detected by two independent anti-KEAP1 antibodies that recognize different epitopes, 7045 against C-terminus domain and 22-002 against recombinant fusion protein of human KEAP1.  KEAP1 proteins are detected in all the tested cell lines at different expression levels by the two independent antibodies.  

KO Validation (Figure 3): Anti-KEAP1 antibodies (7045) specificity was further verified by KEAP1 specific knockout. KEAP1 signal was not detected in KEAP1 knockout HeLa cells as compared to that in control wild type cells.

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