Overview

  • 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.
  • 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

  • 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...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR22664-26
  • Isotype

    IgG
  • Research areas

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
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.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IP.
  • Target

    • 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
      modifications

      Ubiquitinated and subject to proteasomal degradation.
    • Cellular localization

      Cytoplasm. 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

    Images

    • 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 minutes


      Blocking/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.

    • 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).

    • 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).

    References

    ab227828 has not yet been referenced specifically in any publications.

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