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Validated using a knockout cell line

Anti-PDK1 antibody [2H3AA11] (ab110335)

Submit a review Submit a question References (5)
Western blot - Anti-PDK1 antibody [2H3AA11] (ab110335)
  • Western blot - Anti-PDK1 antibody [2H3AA11] (ab110335)
  • Western blot - Anti-PDK1 antibody [2H3AA11] (ab110335)
  • Flow Cytometry - Anti-PDK1 antibody [2H3AA11] (ab110335)
  • Immunocytochemistry/ Immunofluorescence - Anti-PDK1 antibody [2H3AA11] (ab110335)

Key features and details

  • Mouse monoclonal [2H3AA11] to PDK1
  • Suitable for: ICC/IF, WB, Flow Cyt
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-PDK1 antibody [2H3AA11]
    See all PDK1 primary antibodies

  • Description

    Mouse monoclonal [2H3AA11] to PDK1

  • Host species

    Mouse

  • Specificity

    This antibody does not cross-react with other isoforms of human PDK (i.e. PDK2, 3 or 4) and does not cross-react with rat or mouse samples.

  • Tested applications

    Suitable for: ICC/IF, WB, Flow Cytmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • HeLa cells; Human heart and liver tissue lysates; and HL-60 cells

  • General notes

    This monoclonal antibody to PDK1 has been knockout validated in Western blot. The expected band for PDK1 was observed in wild type cells and the band was not seen in knockout cells.

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    Product was previously marketed under the MitoSciences sub-brand.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab110335 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 1 µg/ml.
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 49 kDa.
Flow Cyt
Use a concentration of 1 µg/ml.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Notes
ICC/IF
Use a concentration of 1 µg/ml.
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 49 kDa.
Flow Cyt
Use a concentration of 1 µg/ml.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

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

    • 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 [2H3AA11] (ab110335)
      Western blot - Anti-PDK1 antibody [2H3AA11] (ab110335)

      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 - ab110335 observed at 46 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab110335 detected the expected band for PDK1 in wild-type cells and the band was not seen in knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and PDK1 knockout samples were subjected to SDS-PAGE. ab110335 and ab8245 (loading control to GAPDH) were diluted 1 μg/mL and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10 000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-PDK1 antibody [2H3AA11] (ab110335)
      Western blot - Anti-PDK1 antibody [2H3AA11] (ab110335)
      All lanes : Anti-PDK1 antibody [2H3AA11] (ab110335) at 1 µg/ml

      Lane 1 : Human heart lysate at 10 µg
      Lane 2 : Human liver lysate at 10 µg
      Lane 3 : Recombinant Human PDK1 at 0.01 µg
      Lane 4 : Recombinant Human PDK2 at 0.01 µg
      Lane 5 : Recombinant Human PDK3 at 0.01 µg
      Lane 6 : Recombinant Human PDK4 at 0.01 µg

      Predicted band size: 49 kDa

    • Western blot - Anti-PDK1 antibody [2H3AA11] (ab110335)
      Western blot - Anti-PDK1 antibody [2H3AA11] (ab110335)
      All lanes : Anti-PDK1 antibody [2H3AA11] (ab110335) at 1 µg/ml

      Lane 1 : HeLa cells incubated for 24 hours with vehicle
      Lane 2 : HeLa cells incubated for 24 hours with 100 µM cobalt chloride (CoCl2)
      Lane 3 : HeLa cells incubated for 24 hours with 1 mM deferoxamine (DFO)

      Lysates/proteins at 30 µg per lane.

      Predicted band size: 49 kDa



      Treatment with either CoCl2 or DFO results in increased expression of PDK1, which is known to be transcriptionally upregulated in response to hypoxia.
    • Flow Cytometry - Anti-PDK1 antibody [2H3AA11] (ab110335)
      Flow Cytometry - Anti-PDK1 antibody [2H3AA11] (ab110335)
      HL-60 cells were stained with 1 µg/mL PDK1 antibody (ab110335) (blue) or an equal amount of an isotype control antibody (red) and analyzed by flow cytometry.
    • Immunocytochemistry/ Immunofluorescence - Anti-PDK1 antibody [2H3AA11] (ab110335)
      Immunocytochemistry/ Immunofluorescence - Anti-PDK1 antibody [2H3AA11] (ab110335)
      Immunocytochemistry image of PDK1 stained HeLa cells. The Upper image shows cells which were paraformaldehyde fixed (4%, 20 minutes) and Triton X-100 permeabilized (0.1%, 15 minutes). The cells were incubated with the PDK1 antibody (ab110335) at 2 µg/mL for 2 hours at room temperature or over night at 4°C. The secondary antibody was (green) Alexa Fluor® 488 goat anti-mouse IgG (H+L) at a 1/1000 dilution for 1 hour. 10% Goat serum was used as the blocking agent for all blocking steps. The target protein locates to the mitochondrial matrix.
      The lower image shows cells co-stained with an antibody against HSP60 (red), an enzyme also located in the mitochondrial matrix. The composite image shows an identical mitochondrial pattern for both antibodies indicated by merged orange color.

    References (5)

    Publishing research using ab110335? Please let us know so that we can cite the reference in this datasheet.

    ab110335 has been referenced in 5 publications.

    • Sun S  et al. MicroRNA let-7i-3p affects osteoblast differentiation in ankylosing spondylitis via targeting PDK1. Cell Cycle 20:1209-1219 (2021). PubMed: 34048311
    • Nguyen QL  et al. Alterations in platelet bioenergetics in Group 2 PH-HFpEF patients. PLoS One 14:e0220490 (2019). PubMed: 31365585
    • Nguyen QL  et al. Platelets from pulmonary hypertension patients show increased mitochondrial reserve capacity. JCI Insight 2:e91415 (2017). PubMed: 28289721
    • Ahn E  et al. Temporal fluxomics reveals oscillations in TCA cycle flux throughout the mammalian cell cycle. Mol Syst Biol 13:953 (2017). PubMed: 29109155
    • Goto M  et al. Adaptation of leukemia cells to hypoxic condition through switching the energy metabolism or avoiding the oxidative stress. BMC Cancer 14:76 (2014). WB ; Human . PubMed: 24506813

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