Human Pyruvate Dehydrogenase Kinase isoform 4 ELISA Kit (ab126582)

Overview

  • Product name
    Human Pyruvate Dehydrogenase Kinase isoform 4 ELISA Kit
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    PDK4 3 4%
    Heart Lysate 3 6%
    Inter-assay
    Sample n Mean SD CV%
    PDK4 3 0.7%
    Heart Lysate 3 1.5%
  • Sample type
    Tissue Extracts
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    < 3.9 ng/ml
  • Range
    3.9 ng/ml - 250 ng/ml
  • Assay duration
    Multiple steps standard assay
  • Species reactivity
    Reacts with: Human
  • Product overview

    The oxidation of carbohydrates in mammals is regulated by the pyruvate dehydrogenase (PDH) complex, which is covalently regulated by four PDH kinases (PDK1-4) and two PDH phosphatases (PDP1-2) through a phosphorylation-dephosphorylation cycle. PDK4 Inhibits the mitochondrial pyruvate dehydrogenase complex by phosphorylation of the E1 alpha subunit, thus contributing to the regulation of glucose metabolism. PDK4 exists ubiquitous with highest levels of expression in heart and skeletal muscle. PDK4 transcription and PDK4 mRNA are markedly increased in human skeletal muscle during prolonged exercise and after both short-term high-intensity and prolonged low-intensity exercise. The exercise-induced transcriptional response of PDK4 is enhanced when muscle glycogen is lowered before the exercise, and intake of a low-carbohydrate high-fat diet during recovery from exercise results in increased transcription and mRNA content of PDK4 when compared with intake of a high-carbohydrate diet. PDK4 expression is also increased in response to fasting and a high-fat diet.

    ab126582 PDK4 Human ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the accurate quantitative measurement of PDK4 protein in human tissue lysates. The assay employs an antibody specific to PDK4 protein coated onto well plate strips. Standards and samples are pipetted into the wells and PDK4 present in the sample is bound to the wells by the immobilized antibody. The wells are washed and a primary anti-PDK4 detector antibody is added. After washing away unbound detector antibody, HRP-conjugated secondary antibody (HRP Label) specific for the primary detector antibody is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of bound PDK4. The developing blue color is measured at 600 nm. Optionally the reaction can be stopped by adding hydrochloric acid which changes the color from blue to yellow and the intensity can be measured at 450 nm.

  • Tested applications
    Suitable for: ELISAmore details
  • Platform
    Microplate

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab126582 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
ELISA Use at an assay dependent concentration.

Images

  • Example standard curve of serially diluted purified human PDK4 in the linear working range of the assay.
  • Example human sample curve of serially diluted human heart homogenate extract in the working range of the assay.
  • Interference by cell culture media (20F DMEM), Goat serum, and a titration of 1X extraction buffer were evaluated.
  • PDK4 exists ubiquitous in tissues with highest levels of expression in heart (HHH) and skeletal muscle (HSMH). This ELISA assay applies specifically to human samples. No signal was observed when using rat tissue (rat heart homogenate (RHH)) or mouse tissue (mouse heart homogenate (MHH)).
  • Assay Specificity was determined by using human PDK1 recombinant protein as negative control.
  • All lanes : Human Pyruvate Dehydrogenase Kinase isoform 4 ELISA Kit (ab126582)

    Lane 1 : HepG2 whole cell lysate (ab7900) at 20 µg
    Lane 2 : Human heart tissue lysate - total protein (ab29431) at 20 µg
    Lane 3 : PDK1 protein at 0.1 µg
    Lane 4 : PDK2 protein at 0.1 µg
    Lane 5 : PDK3 protein at 0.1 µg
    Lane 6 : PDK4 protein at 0.1 µg

Protocols

References

ab126582 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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