

Recombinant Anti-Keap1 antibody [EPR22664-26] (ab227828)
- Datasheet
- References
- Protocols
Overview
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Product name
Anti-Keap1 antibody [EPR22664-26]
See all Keap1 primary antibodies -
Description
Rabbit monoclonal [EPR22664-26] to Keap1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Recombinant fragment within Human Keap1 aa 300 to the C-terminus. The exact sequence is proprietary.
Database link: Q14145 -
Positive control
- WB: Human lung tissue lysate; Wild-type HAP1, NK-92 and HepG2 whole cell lysates; mouse E14.5 liver tissue lysate. IHC-P: Human lung carcinoma tissue; mouse kidney tissue.
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General notes
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
This product is a recombinant rabbit monoclonal antibody.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22664-26 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
Our Abpromise guarantee covers the use of ab227828 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB | 1/1000. Detects a band of approximately 70 kDa (predicted molecular weight: 70 kDa). | |
IHC-P | 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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Function
Retains NFE2L2/NRF2 in the cytosol. Functions as substrate adapter protein for the E3 ubiquitin ligase complex formed by CUL3 and RBX1. Targets NFE2L2/NRF2 for ubiquitination and degradation by the proteasome, thus resulting in the suppression of its transcriptional activity and the repression of antioxidant response element-mediated detoxifying enzyme gene expression. May also retain BPTF in the cytosol. Targets PGAM5 for ubiquitination and degradation by the proteasome. -
Tissue specificity
Broadly expressed, with highest levels in skeletal muscle. -
Sequence similarities
Contains 1 BACK (BTB/Kelch associated) domain.
Contains 1 BTB (POZ) domain.
Contains 6 Kelch repeats. -
Domain
The Kelch repeats mediate interaction with NF2L2/NRF2, BPTF and PGAM5. -
Post-translational
modificationsUbiquitinated and subject to proteasomal degradation. -
Cellular localization
Cytoplasm. Nucleus. Shuttles between cytoplasm and nucleus. - Information by UniProt
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Database links
- Entrez Gene: 9817 Human
- Entrez Gene: 50868 Mouse
- Omim: 606016 Human
- SwissProt: Q14145 Human
- SwissProt: Q9Z2X8 Mouse
- Unigene: 465870 Human
- Unigene: 248266 Mouse
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Alternative names
- Cytosolic inhibitor of Nrf2 antibody
- INrf 2 antibody
- INrf2 antibody
see all
Images
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All lanes : Anti-Keap1 antibody [EPR22664-26] (ab227828) at 1/1000 dilution
Lane 1 : Human lung tissue lysate
Lane 2 : Wild-type HAP1 whole cell lysate
Lane 3 : Keap1 knockout HAP1 whole cell lysate
Lane 4 : NK-92 (human malignant non-Hodgkin's lymphoma natural killer cell), whole cell lysate
Lane 5 : HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate
Lane 6 : Mouse E14.5 liver tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
Lane 1 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Lanes 2-6 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 70 kDa
Observed band size: 70 kDa
Exposure time: 3 minutesBlocking/diluting buffer and concentration: 5% NFDM/TBST
ab227828 was shown to specifically react with Keap1 in wild-type HAP1 cells as signal was lost in Keap1 knockout cells. Wild-type and Keap1 knockout samples were subjected to SDS-PAGE. ab227828 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 23242280, PMID: 20404090).
This blot was developed using a higher sensitivity ECL substrate.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Keap1 antibody [EPR22664-26] (ab227828)
Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Keap1 with ab227828 at 1/500 dilution (0.97 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on kidney tubules of mouse kidney (PMID: 28324005) is observed. The section was incubated with ab227828 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Keap1 antibody [EPR22664-26] (ab227828)
Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labeling Keap1 with ab227828 at 1/500 dilution (0.97 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on tumor cells of human lung carcinoma (PMID: 20534738) is observed. The section was incubated with ab227828 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Datasheets and documents
References
ab227828 has not yet been referenced specifically in any publications.