Overview

  • Product nameAnti-Keap1 antibody
    See all Keap1 primary antibodies
  • Description
    Rabbit polyclonal to Keap1
  • Tested applicationsSuitable for: ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Rabbit, Horse, Guinea pig, Cow, Cat, Dog, Pig, Zebrafish
  • Immunogen

    Synthetic peptide: ETWTFVAPMK HRRSALGITV HQGRIYVLGG YDGHTFLDSV ECYDPDTDTW , corresponding to amino acids 542-591 of Human Keap1

  • Positive control
    • HepG2 cell lysate. Human kidney tissue.
  • General notesExpression of Keap1 shuttles between cytoplasm and nucleus.

Properties

Applications

Our Abpromise guarantee covers the use of ab66620 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
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 70 kDa (predicted molecular weight: 70 kDa).Can be blocked with Human Keap1 peptide (ab111654). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
IHC-P Use a concentration of 4 - 8 µg/ml.

Target

  • FunctionRetains 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 specificityBroadly expressed, with highest levels in skeletal muscle.
  • Sequence similaritiesContains 1 BACK (BTB/Kelch associated) domain.
    Contains 1 BTB (POZ) domain.
    Contains 6 Kelch repeats.
  • DomainThe Kelch repeats mediate interaction with NF2L2/NRF2, BPTF and PGAM5.
  • Post-translational
    modifications
    Ubiquitinated and subject to proteasomal degradation.
  • Cellular localizationCytoplasm. Nucleus. Shuttles between cytoplasm and nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Cytosolic inhibitor of Nrf2 antibody
    • INrf 2 antibody
    • INrf2 antibody
    • Keap 1 antibody
    • KEAP1 antibody
    • KEAP1_HUMAN antibody
    • Kelch like ECH associated protein 1 antibody
    • Kelch like family member 19 antibody
    • Kelch like protein 19 antibody
    • Kelch-like ECH-associated protein 1 antibody
    • Kelch-like protein 19 antibody
    • KIAA0132 antibody
    • KLHL 19 antibody
    • KLHL19 antibody
    • MGC10630 antibody
    • MGC1114 antibody
    • MGC20887 antibody
    • MGC4407 antibody
    • MGC9454 antibody
    see all

Anti-Keap1 antibody images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling Keap1 with ab66620 at a concentration of 4-8µg/ml. Arrows indicate positively stained epithelial cells of the renal tubule. Magnification: 400X.

  • ICC/IF image of ab66620 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab66620, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Lane 1 : Anti-Keap1 antibody (ab66620) at 5 µg/ml
    Lane 2 : Anti-Keap1 antibody (ab66620) at 2.5 µg/ml
    Lane 3 : Anti-Keap1 antibody (ab66620) at 1.25 µg/ml

    Lane 1 : Cell lysate prepared form HepG2 cells
    Lane 2 : Cell lysate prepared form HepG2 cells
    Lane 3 : Cell lysate prepared form HepG2 cells

    Lysates/proteins at 25 µg/ml per lane.


    Predicted band size : 70 kDa
  • All lanes : Anti-Keap1 antibody (ab66620) at 1.25 µg/ml

    Lane 1 : HepG2 cell lysate
    Lane 2 : HepG2 cell lysate with blocking peptide at 1 µg/ml

    Lysates/proteins at 25 µg per lane.


    Predicted band size : 70 kDa
  • Anti-Keap1 antibody (ab66620) at 1 µg/ml + Lysate prepared from human 293T cells at 10 µg

    Secondary
    IRDye® 700DX-conjugated donkey polyclonal to rabbit IgG at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 70 kDa
    Observed band size : 70 kDa
    Additional bands at : 36 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 1 minute

    This image is a courtesy of Anonymous Abreview

    See Abreview

References for Anti-Keap1 antibody (ab66620)

This product has been referenced in:
  • Orthwein A  et al. A mechanism for the suppression of homologous recombination in G1 cells. Nature 528:422-6 (2015). Read more (PubMed: 26649820) »
  • Baqader NO  et al. Nuclear cytoplasmic trafficking of proteins is a major response of human fibroblasts to oxidative stress. J Proteome Res 13:4398-423 (2014). WB ; Human . Read more (PubMed: 25133973) »

See all 4 Publications for this product

Product Wall

Application Western blot
Sample Dog Tissue lysate - other (brain)
Gel Running Conditions Reduced Denaturing (4-20%)
Loading amount 20 µg
Specification brain
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 24°C
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Submitted Aug 19 2016

Application Western blot
Sample Rat Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions Reduced Denaturing (10%)
Loading amount 20 µg
Specification Skeletal muscle
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
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Submitted Feb 29 2016

Application Western blot
Sample Mouse Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions Reduced Denaturing (10%)
Loading amount 20 µg
Specification Skeletal muscle
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
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Submitted Feb 29 2016

Application Western blot
Sample Mouse Tissue lysate - nuclear (liver nuclear fraction)
Gel Running Conditions Reduced Denaturing (10% gel)
Loading amount 35 µg
Treatment hyperoxia for 44 hours
Specification liver nuclear fraction
Blocking step Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Dr. Sandra Sobocanec

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Submitted Nov 19 2015

Application Western blot
Loading amount 15 µg
Gel Running Conditions Reduced Denaturing (10% gel)
Sample Human Cell lysate - whole cell (Jurkat cell lysate)
Specification Jurkat cell lysate
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
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Submitted Jun 02 2014

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (10% gel)
Sample Mouse Tissue lysate - whole (Skeletal muscle)
Specification Skeletal muscle
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
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Submitted Jun 02 2014

Gracias por contactarnos de nuevo.

El anticuerpo Anti-Caveolin 1 (ab18199) ha sido testado en inmunoprecipitación, Western Blot e immunohistoquímica en secciones embebidas en parafina. Al haberse validado en estas aplicaciones e...

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Gracias por contactarnos.

El producto ab66620 tiene un precio de €380.00 y actualmente en nuestra oficina de Inglaterra queda una unidad es stock. Los productos ab31163 y ab18199 valen €410.00 cada uno y tenemos suficientes unidades...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (THP-1)
Loading amount 20 µg
Specification THP-1
Gel Running Conditions Reduced Denaturing (5-20% gradient)
Blocking step Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5%
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Submitted Jan 07 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (293T)
Loading amount 10 µg
Specification 293T
Gel Running Conditions Reduced Denaturing (4-12%)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Oct 26 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"