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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).
Figure 1 Western Blot Validation of KEAP1 in Human Lung
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.
Cell Treatment
IAV
KO
Trial Size

KEAP1 Antibody

CATALOG NUMBER: 7045

Clonality:
Polyclonal
Tested Applications:
ELISA, IF, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
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.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IF, WB
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.
Specificity:
At least two isoforms of KEAP1 are known to exist.
Positive Control 1:
Cat. No. 1302 - Human Lung Tissue Lysate
Positive Control 2:
Cat. No. 10-101 - Human Lung Tissue Slide
Positive Control 3:
Cat. No. 1224 - Daudi Cell Lysate
Positive Control 4:
Cat. No. 1204– K562 Cell Lysate
Predicted Molecular Weight:
Predicted: 70kD
Observed: 65-70kD
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.

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.
Purification:
KEAP1 Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
KEAP1 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
KEAP1 antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
KEAP1
Additional Names:
KEAP1 Antibody: INrf2, KLHL19, INRF2, KIAA0132, Kelch-like ECH-associated protein 1, Cytosolic inhibitor of Nrf2, INrf2
Protein Accession Number:
NP_987096
Protein Gi Number:
45269145
Ncbi Gene Id Number:
9817
User Note:
Optimal dilutions for each application to be determined by the researcher.
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 Reference 1:
Zhang et al. Mol. Cell. Biol. 2004;24:10941-53.
Background Reference 2:
Kobayashi et al. Mol. Cell. Biol. 2004; 24:7130-9.
Background Reference 3:
Jiang et al. Int. J. Mol. Med. 2012; 29:946-56.
Background Reference 4:
Devling et al. Proc. Natl. Acad. Sci. USA 2005; 102: 7280-5A.

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

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.

CATALOG NUMBER:

7045

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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