Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-PDK1 antibody [EPR19571] (ab202468)

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Western blot - Anti-PDK1 antibody [EPR19571] (ab202468)
  • Immunocytochemistry/ Immunofluorescence - Anti-PDK1 antibody [EPR19571] (ab202468)
  • Flow Cytometry - Anti-PDK1 antibody [EPR19571] (ab202468)
  • Immunoprecipitation - Anti-PDK1 antibody [EPR19571] (ab202468)
  • Western blot - Anti-PDK1 antibody [EPR19571] (ab202468)
  • Immunocytochemistry/ Immunofluorescence - Anti-PDK1 antibody [EPR19571] (ab202468)
  • Western blot - Anti-PDK1 antibody [EPR19571] (ab202468)
  • Anti-PDK1 antibody [EPR19571] (ab202468)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19571] to PDK1
  • Suitable for: WB, ICC/IF, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-PDK1 antibody [EPR19571]
    See all PDK1 primary antibodies

  • Description

    Rabbit monoclonal [EPR19571] to PDK1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: L-929, NIH/3T3, F9, bEnd.3, C6 and PC-12 whole cell lysates; Mouse brain, kidney, spleen and heart lysates; Rat brain, kidney and heart lysates. ICC/IF: NIH/3T3 and RAW 264.7 cells. Flow Cyt: NIH/3T3 cells. IP: NIH/3T3 whole cell 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

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab202468 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
1/2000. Detects a band of approximately 47 kDa (predicted molecular weight: 49 kDa).
ICC/IF
1/1000.
Flow Cyt
1/600.
IP
1/40.
Notes
WB
1/2000. Detects a band of approximately 47 kDa (predicted molecular weight: 49 kDa).
ICC/IF
1/1000.
Flow Cyt
1/600.
IP
1/40.

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 [EPR19571] (ab202468)
    Western blot - Anti-PDK1 antibody [EPR19571] (ab202468)

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: PDK1 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: Jurkat whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab202468 observed at 45 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab202468 was shown to specifically react with PDK1 in wild-type cells along with additional cross reactive bands. Signal was lost in PDK1 knockout cells. Wild-type and PDK1 knockout samples were subjected to SDS-PAGE. ab202468 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-PDK1 antibody [EPR19571] (ab202468)
    Immunocytochemistry/ Immunofluorescence - Anti-PDK1 antibody [EPR19571] (ab202468)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling PDK1 with ab202468 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasmic staining on NIH/3T3 cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab202468 at 1/1000 dilution followed by ab150120  at 1/1000 dilution.

    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

  • Flow Cytometry - Anti-PDK1 antibody [EPR19571] (ab202468)
    Flow Cytometry - Anti-PDK1 antibody [EPR19571] (ab202468)

    Flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling PDK1 with ab202468 at 1/600 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.

    Cells were permeabilised by 90% methanol-1XPBS, -20ºC, 30min.

  • Immunoprecipitation - Anti-PDK1 antibody [EPR19571] (ab202468)
    Immunoprecipitation - Anti-PDK1 antibody [EPR19571] (ab202468)

    PDK1 was immunoprecipitated from 1mg of NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate with ab202468 at 1/40 dilution.

    Western blot was performed from the immunoprecipitate using ab202468 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: NIH/3T3 whole cell lysate, 10µg (Input).

    Lane 2: ab202468 IP in NIH/3T3 whole cell lysate.

    Lane 3:  Rabbit IgG,monoclonal -[EPR25A] - Isotype Control (ab172730) instead of ab202468 in NIH/3T3 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 1 second.

  • Western blot - Anti-PDK1 antibody [EPR19571] (ab202468)
    Western blot - Anti-PDK1 antibody [EPR19571] (ab202468)
    All lanes : Anti-PDK1 antibody [EPR19571] (ab202468) at 1/2000 dilution

    Lane 1 : L-929 (Mouse connective tissue fibroblast cell line) whole cell lysate
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
    Lane 3 : F9 (Mouse embryonic testicular cancer cell line) whole cell lysate
    Lane 4 : bEnd.3 (Mouse brain endothelioma cell line) whole cell lysate
    Lane 5 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 6 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 49 kDa
    Observed band size: 47 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 1 minute; Lane 3-4: 10 seconds; Lane 5/6: 8 seconds.

  • Immunocytochemistry/ Immunofluorescence - Anti-PDK1 antibody [EPR19571] (ab202468)
    Immunocytochemistry/ Immunofluorescence - Anti-PDK1 antibody [EPR19571] (ab202468)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling PDK1 with ab202468 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasmic staining on RAW 264.7 cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab202468 at 1/1000 dilution followed by ab150120 at 1/1000 dilution.

    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

  • Western blot - Anti-PDK1 antibody [EPR19571] (ab202468)
    Western blot - Anti-PDK1 antibody [EPR19571] (ab202468)
    All lanes : Anti-PDK1 antibody [EPR19571] (ab202468) at 1/2000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse kidney lysate
    Lane 3 : Mouse spleen lysate
    Lane 4 : Mouse heart lysate
    Lane 5 : Rat brain lysate
    Lane 6 : Rat kidney lysate
    Lane 7 : Rat heart lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 49 kDa
    Observed band size: 47 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1/2/3/5/6: 8 seconds; Lane 4: 3 seconds; Lane 7: 1 second.

  • Anti-PDK1 antibody [EPR19571] (ab202468)
    Anti-PDK1 antibody [EPR19571] (ab202468)

References (2)

Publishing research using ab202468? Please let us know so that we can cite the reference in this datasheet.

ab202468 has been referenced in 2 publications.

  • Xu H  et al. Inhibition of pyruvate dehydrogenase kinase-1 by dicoumarol enhances the sensitivity of hepatocellular carcinoma cells to oxaliplatin via metabolic reprogramming. Int J Oncol 57:733-742 (2020). PubMed: 32705170
  • He L  et al. Puerarin inhibits apoptosis and inflammation in myocardial cells via PPARa expression in rats with chronic heart failure. Exp Ther Med 18:3347-3356 (2019). PubMed: 31602208

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