Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-PDK1 antibody [EPR13009] (ab175190)

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Overview

  • Product name

    Anti-PDK1 antibody [EPR13009]
    See all PDK1 primary antibodies

  • Description

    Rabbit monoclonal [EPR13009] to PDK1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IPmore details
    Unsuitable for: Flow Cyt,ICC/IF or IHC-P

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human PDK1 aa 1-100 (Cysteine residue). The exact sequence is proprietary.
    Database link: Q15118

  • Positive control

    • WB: Hap1, HeLa, LNCaP and Human fetal heart lysate. IP: Human fetal heart lysate.

  • General notes

     This product was previously labelled as Mitochondrial Pyruvate dehydrogenase kinase 1

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

Our Abpromise guarantee covers the use of ab175190 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 - 1/10000. Predicted molecular weight: 49 kDa.
IP 1/10 - 1/100.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IHC-P.

    • Target

    • Function

      Inhibits the mitochondrial pyruvate dehydrogenase complex by phosphorylation of the E1 alpha subunit, thus contributing to the regulation of glucose metabolism.

    • Tissue specificity

      Expressed predominantly in the heart.

    • Sequence similarities

      Belongs to the PDK/BCKDK protein kinase family.
      Contains 1 histidine kinase domain.

    • Cellular localization

      Mitochondrion matrix.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

      • [Pyruvate dehydrogenase [lipoamide]] kinase isozyme 1, mitochondrial antibody
      • HGNC:8809 antibody
      • Mitochondrial pyruvate dehydrogenase kinase isoenzyme 1 antibody
      • PDH kinase 1 antibody
      • Pdk1 antibody
      • PDK1_HUMAN antibody
      • Pyruvate dehydrogenase kinase isoform 1 antibody
      • Pyruvate dehydrogenase kinase, isoenzyme 1 antibody
      see all

    Images

    • Western blot - Anti-PDK1 antibody [EPR13009] (ab175190)
      Western blot - Anti-PDK1 antibody [EPR13009] (ab175190)
      All lanes : Anti-PDK1 antibody [EPR13009] (ab175190) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : PDK1 knockout HeLa cell lysate
      Lane 3 : Wild-type HAP1 cell lysate
      Lane 4 : PDK1 knockout HAP1 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 49 kDa



      Lanes 1- 4: Merged signal (red and green). Green - ab175190 observed at 46 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

      ab175190 was shown to react with PDK1 in wild-type HeLa cells in Western blot. Loss of signal was observed when knockout sample ab257164 was used. Wild-type HeLa and PDK1 knockout HeLa whole cell lysates were subjected to SDS-PAGE. ab175190 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-PDK1 antibody [EPR13009] (ab175190)
      Western blot - Anti-PDK1 antibody [EPR13009] (ab175190)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: PDK1 knockout HAP1 cell lysate (20 µg)
      Lane 3: Human fetal heart lysate (20 µg)
      Lane 4: LnCaP cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab175190 observed at 46 kDa. Red - loading control, ab8245, observed at 37 kDa.


      ab175190 was shown to specifically recognize PDK1 in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when PDK1 knockout samples were examined. Wild-type and PDK1 knockout samples were subjected to SDS-PAGE. ab175190 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-PDK1 antibody [EPR13009] (ab175190)
      Western blot - Anti-PDK1 antibody [EPR13009] (ab175190)
      All lanes : Anti-PDK1 antibody [EPR13009] (ab175190) at 1/1000 dilution

      Lane 1 : LNCaP lysates
      Lane 2 : Human fetal heart lysates

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 49 kDa

    • Immunoprecipitation - Anti-PDK1 antibody [EPR13009] (ab175190)
      Immunoprecipitation - Anti-PDK1 antibody [EPR13009] (ab175190)

      Western blot analysis on immunoprecipitation pellet from Human fetal heart lysate (lane 1) labeling PDK1 with ab175190 at 1/10 dilution or 1X PBS (lane 2) negative control.  HRP-conjugated anti-Rabbit IgG preferentially detecting the non-reduced form of Rabbit IgG.

    References

    This product has been referenced in:

    • Wang F  et al. Hypoxia regulates sumoylation pathways in intervertebral disc cells: implications for hypoxic adaptations. Osteoarthritis Cartilage 24:1113-24 (2016). Read more (PubMed: 26826302) »
    See 1 Publication for this product

    Publishing research using ab175190? Please let us know so that we can cite the reference in this datasheet.

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