Overview

  • Product name

    Anti-PDK1 antibody [EPR19573]
    See all PDK1 primary antibodies
  • Description

    Rabbit monoclonal [EPR19573] to PDK1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human PDK1 aa 150 to the C-terminus. The exact sequence is proprietary.
    Database link: Q15118

  • Positive control

    • WB: HEK-293, Jurkat, HepG2, NIH/3T3, PC-12 and LNCaP whole cell lysates; human heart and kidney lysates; mouse brain, heart, kidney and spleen lysates; rat kidney and spleen lysates; HEK-293 mitochondria fraction lysate. IP: HepG2 whole cell lysate.
  • General notes

     

     

     This product was previously labelled as Mitochondrial Pyruvate dehydrogenase kinase 1

     

    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

Applications

Our Abpromise guarantee covers the use of ab207450 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).
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

  • 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

  • Lane 1: Wild-type HAP1 cell lysate (40 µg)
    Lane 2: PDK1 knockout HAP1 cell lysate (40 µg)
    Lane 3: HepG2 cell lysate (20 µg)
    Lane 4: LnCaP cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab207450 observed at 45 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab207450 was shown to specifically react with PDK1 in wild-type HAP1 cells. No band was observed when PDK1 knockout samples were examined. Wild-type and PDK1 knockout samples were subjected to SDS-PAGE. Ab207450 and ab8245 (loading control to GAPDH) were diluted at 1/2000 and 1/10,000 dilution 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 1/10,000 dilution for 1 hour at room temperature before imaging.

  • PDK1 was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate with ab207450 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab207450 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HepG2 whole cell lysate 10µg (Input).

    Lane 2: ab207450 IP in HepG2 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207450 in HepG2 whole cell lysate.

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

    Exposure time: 1 second.

  • All lanes : Anti-PDK1 antibody [EPR19573] (ab207450) at 1/2000 dilution

    Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 3 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
    Lane 5 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 6 : LNCaP (Human prostate cancer cell line) whole cell 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 1-5: 15 seconds; Lane 6: 8 seconds.

     

  • All lanes : Anti-PDK1 antibody [EPR19573] (ab207450) at 1/2000 dilution

    Lane 1 : Human heart lysate
    Lane 2 : Human kidney lysate
    Lane 3 : Mouse brain lysate
    Lane 4 : Mouse heart lysate
    Lane 5 : Mouse kidney lysate
    Lane 6 : Mouse spleen lysate
    Lane 7 : Rat kidney lysate
    Lane 8 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lanes 1-2 : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
    Lanes 3-8 : 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 1/3/4: 8 seconds; Lane 2: 30 seconds; Lane 5/6: 3 minutes; Lane 7/8: 15 seconds.

  • All lanes : Anti-PDK1 antibody [EPR19573] (ab207450) at 1/5000 dilution

    Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) mitochondria fraction lysate
    Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) cytosol fraction 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?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

References

This product has been referenced in:

  • Raghubeer S  et al. Ochratoxin A upregulates biomarkers associated with hypoxia and transformation in human kidney cells. Toxicol In Vitro 57:211-216 (2019). Read more (PubMed: 30876885) »
  • Sikorski K  et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). Read more (PubMed: 30377371) »
See all 4 Publications for this product

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