RecombinantRabMAb

Recombinant Anti-PDK4 antibody [EPR19727-245] (ab214938)

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Western blot - Anti-PDK4 antibody [EPR19727-245] (ab214938)
  • Western blot - Anti-PDK4 antibody [EPR19727-245] (ab214938)
  • Western blot - Anti-PDK4 antibody [EPR19727-245] (ab214938)
  • Flow Cytometry (Intracellular) - Anti-PDK4 antibody [EPR19727-245] (ab214938)
  • Immunoprecipitation - Anti-PDK4 antibody [EPR19727-245] (ab214938)
  • Anti-PDK4 antibody [EPR19727-245] (ab214938)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19727-245] to PDK4
  • Suitable for: IP, WB, Flow Cyt (Intra)
  • Reacts with: Mouse, Rat, Recombinant fragment

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-PDK4 antibody [EPR19727-245]
    See all PDK4 primary antibodies

  • Description

    Rabbit monoclonal [EPR19727-245] to PDK4

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, WB, Flow Cyt (Intra)more details

  • Species reactivity

    Reacts with: Mouse, Rat, Recombinant fragment

  • Immunogen

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

  • Positive control

    • WB: Mouse PDK4 recombinant protein; mouse skeletal muscle and heart lysates; rat muscle and heart lysates; C2C12 and C6 whole cell lysate; heart from nonfasting mouse and mouse fasting for 72 hours; gastrocnemius muscle from nonfasting mouse and mouse fasting for 72 hours. Flow Cyt (intra): C2C12 cells. IP: Mouse heart 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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab214938 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
IP
1/30.
WB
1/1000. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
Flow Cyt (Intra)
1/60.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Notes
IP
1/30.
WB
1/1000. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
Flow Cyt (Intra)
1/60.

ab172730 - Rabbit monoclonal IgG, 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

    Ubiquitous; highest levels of expression in heart and skeletal muscle.

  • 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 4 antibody
      • FLJ40832 antibody
      • mitochondrial antibody
      • Pdk4 antibody
      • PDK4_HUMAN antibody
      • Pyruvate dehydrogenase [lipoamide] kinase isozyme 4 mitochondrial antibody
      • Pyruvate dehydrogenase kinase 4 antibody
      • Pyruvate dehydrogenase kinase isoenzyme 4 antibody
      • Pyruvate dehydrogenase kinase isoform 4 antibody
      • Pyruvate dehydrogenase kinase isozyme 4 antibody
      • Pyruvate dehydrogenase kinase isozyme 4 mitochondrial antibody
      see all

    Images

    • Western blot - Anti-PDK4 antibody [EPR19727-245] (ab214938)
      Western blot - Anti-PDK4 antibody [EPR19727-245] (ab214938)
      All lanes : Anti-PDK4 antibody [EPR19727-245] (ab214938) at 1/10000 dilution

      Lane 1 : Mouse PDK4 recombinant protein
      Lane 2 : Mouse PDK1 recombinant protein
      Lane 3 : Mouse PDK2 recombinant protein

      Lysates/proteins at 0.01 µg per lane.

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

      Predicted band size: 47 kDa
      Observed band size: 47 kDa


      Exposure time: 5 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      Mouse PDK4, PDK1 and PDK2 recombinant proteins contain aa8-412, aa1-434 and aa1-407 respectively with a His-Tag®.

    • Western blot - Anti-PDK4 antibody [EPR19727-245] (ab214938)
      Western blot - Anti-PDK4 antibody [EPR19727-245] (ab214938)
      All lanes : Anti-PDK4 antibody [EPR19727-245] (ab214938) at 1/1000 dilution

      Lane 1 : Mouse skeletal muscle lysate
      Lane 2 : Mouse heart lysate
      Lane 3 : Rat muscle lysate
      Lane 4 : Rat heart lysate
      Lane 5 : C2C12 (Mouse myoblast cell line) whole cell lysate
      Lane 6 : C6 (Rat glial tumor 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: 47 kDa
      Additional bands at: 47 kDa. We are unsure as to the identity of these extra bands.



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure time: Lane 1: 5 seconds; Lane 2-4: 3 minutes; Lane 5: 1 minutes; Lane 6: 10 seconds.

    • Western blot - Anti-PDK4 antibody [EPR19727-245] (ab214938)
      Western blot - Anti-PDK4 antibody [EPR19727-245] (ab214938)
      Lanes 1-2 : Anti-PDK4 antibody [EPR19727-245] (ab214938) at 1/10000 dilution
      Lanes 3-4 : Anti-PDK4 antibody [EPR19727-245] (ab214938) at 1/1000 dilution

      Lane 1 : Heart from nonfasting mouse
      Lane 2 : Heart from mouse fasting for 72 hours
      Lane 3 : Gastrocnemius muscle from nonfasting mouse
      Lane 4 : Gastrocnemius muscle from mouse fasting for 72 hours

      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: 47 kDa
      Observed band size: 47 kDa


      Exposure time: 5 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The expression level of PDK4 is increased under the fasting state (PMID: 11723055 and PMID: 16606348).

    • Flow Cytometry (Intracellular) - Anti-PDK4 antibody [EPR19727-245] (ab214938)
      Flow Cytometry (Intracellular) - Anti-PDK4 antibody [EPR19727-245] (ab214938)

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed C2C12 (Mouse myoblast cell line) cells labeling PDK4 with ab214938 at 1/60 dilution (red) compared withRabbit 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 1/2000 dilution was used as the secondary antibody. 

       

       

    • Immunoprecipitation - Anti-PDK4 antibody [EPR19727-245] (ab214938)
      Immunoprecipitation - Anti-PDK4 antibody [EPR19727-245] (ab214938)

      PDK4 was immunoprecipitated from 0.35 mg of mouse heart lysate with ab214938  at 1/30 dilution.

      Western blot was performed from the immunoprecipitate using ab214938 at 1/1000 dilution.

      VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

      Lane 1: Mouse heart lysate 10µg (Input).

      Lane 2: ab214938 IP in mouse heart lysate.

      Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab214938 in mouse heart lysate.

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

      Exposure time: 0.5 second.

    • Anti-PDK4 antibody [EPR19727-245] (ab214938)
      Anti-PDK4 antibody [EPR19727-245] (ab214938)

    References (8)

    Publishing research using ab214938? Please let us know so that we can cite the reference in this datasheet.

    ab214938 has been referenced in 8 publications.

    • Shannon CE  et al. Insulin resistance is mechanistically linked to hepatic mitochondrial remodeling in non-alcoholic fatty liver disease. Mol Metab 45:101154 (2021). PubMed: 33359401
    • Chien HC  et al. PPARd and FOXO1 Mediate Palmitate-Induced Inhibition of Muscle Pyruvate Dehydrogenase Complex and CHO Oxidation, Events Reversed by Electrical Pulse Stimulation. Int J Mol Sci 21:N/A (2020). PubMed: 32824862
    • Hsu JW  et al. Phthalate exposure causes browning-like effects on adipocytes in vitro and in vivo. Food Chem Toxicol 142:111487 (2020). PubMed: 32522588
    • Gao K  et al. Qishen granules exerts cardioprotective effects on rats with heart failure via regulating fatty acid and glucose metabolism. Chin Med 15:21 (2020). PubMed: 32158496
    • Kaya E  et al. Beneficial Effects of Acetyl-DL-Leucine (ADLL) in a Mouse Model of Sandhoff Disease. J Clin Med 9:N/A (2020). PubMed: 32276303
    • Nasci VL  et al. miR-21-5p regulates mitochondrial respiration and lipid content in H9C2 cells. Am J Physiol Heart Circ Physiol 316:H710-H721 (2019). PubMed: 30657727
    • Pin F  et al. PDK4 drives metabolic alterations and muscle atrophy in cancer cachexia. FASEB J 33:7778-7790 (2019). PubMed: 30894018
    • Wu P  et al. MicroRNA-29a is involved lipid metabolism dysfunction and insulin resistance in C2C12 myotubes by targeting PPARd. Mol Med Rep 17:8493-8501 (2018). PubMed: 29693165

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