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    pdk1-antibody-epr19571-ab202468.pdf

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Signal Transduction Metabolism Energy Metabolism
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-PDK1 antibody [EPR19571] (ab202468)

  • Datasheet
  • Certificate of Compliance
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Western blot - Anti-PDK1 antibody [EPR19571] (ab202468)
  • Immunocytochemistry/ Immunofluorescence - Anti-PDK1 antibody [EPR19571] (ab202468)
  • Flow Cytometry (Intracellular) - 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: Flow Cyt (Intra), WB, ICC/IF, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Primary
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Anti-BNIP3 antibody [EPR4034] (ab109362)
Secondary
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Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

View more associated products

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: Flow Cyt (Intra), WB, ICC/IF, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. 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 (intra): NIH/3T3 cells. IP: NIH/3T3 whole cell lysate.
  • General notes

    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

  • 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

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR19571
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Metabolism
    • Energy Metabolism
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of carbohydrates
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Integration of energy metabolism
    • Cancer
    • Cancer Metabolism
    • Cellular metabolic process
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Carbohydrate metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Energy Metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Integration of energy

Associated products

  • Alternative Versions

    • Anti-PDK1 antibody [EPR19571] - BSA and Azide free (ab251373)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (ab210849)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)

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
Flow Cyt (Intra)
1/600.
WB
1/2000. Detects a band of approximately 47 kDa (predicted molecular weight: 49 kDa).
ICC/IF
1/1000.
IP
1/40.
Notes
Flow Cyt (Intra)
1/600.
WB
1/2000. Detects a band of approximately 47 kDa (predicted molecular weight: 49 kDa).
ICC/IF
1/1000.
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.
  • Target information above from: UniProt accession Q15118 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5163 Human
    • Entrez Gene: 228026 Mouse
    • Entrez Gene: 116551 Rat
    • Omim: 602524 Human
    • SwissProt: Q15118 Human
    • SwissProt: Q8BFP9 Mouse
    • SwissProt: Q63065 Rat
    • Unigene: 470633 Human
    • Unigene: 34411 Mouse
    • Unigene: 11185 Rat
    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 (Intracellular) - Anti-PDK1 antibody [EPR19571] (ab202468)
    Flow Cytometry (Intracellular) - Anti-PDK1 antibody [EPR19571] (ab202468)

    Intracellular 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 RabbitIgG,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, -20°C, 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)

Protocols

  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (7)

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

ab202468 has been referenced in 7 publications.

  • He S  et al. Hyperoside protects cardiomyocytes against hypoxia-induced injury via upregulation of microRNA-138. Mol Med Rep 23:N/A (2021). PubMed: 33649812
  • Chen X  et al. Incaspitolide A extracted from Carpesium cernuum induces apoptosis in vitro via the PI3K/AKT pathway in benign prostatic hyperplasia. Biosci Rep 41:N/A (2021). PubMed: 34100062
  • Xu Q  et al. CircHIPK3 regulates pulmonary fibrosis by facilitating glycolysis in miR-30a-3p/FOXK2-dependent manner. Int J Biol Sci 17:2294-2307 (2021). PubMed: 34239356
  • Su X  et al. NOX4-Derived ROS Mediates TGF-ß1-Induced Metabolic Reprogramming during Epithelial-Mesenchymal Transition through the PI3K/AKT/HIF-1a Pathway in Glioblastoma. Oxid Med Cell Longev 2021:5549047 (2021). PubMed: 34257808
  • Chen Z  et al. LncRNA SOX2-OT/miR-30d-5p/PDK1 Regulates PD-L1 Checkpoint Through the mTOR Signaling Pathway to Promote Non-small Cell Lung Cancer Progression and Immune Escape. Front Genet 12:674856 (2021). PubMed: 34394184
  • 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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