Overview

  • Product name

    Mouse PDK4 ELISA Kit
    See all PDK4 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 5 2%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 9%
  • Sample type

    Cell culture extracts, Tissue Extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    45 pg/ml
  • Range

    0.234 ng/ml - 15 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture extracts 98 94% - 103%
    Mouse muscle extract 98 95% - 100%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse, Rat
    Does not react with: Cow, Human
  • Product overview

    PDK4 in vitro SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of PDK4 protein in mouse cell and tissue extract samples.


    The SimpleStep ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    PDK4 is a mitochondrial kinase that plays a key role in regulation of glucose and fatty acid metabolism and homeostasis via phosphorylation of the pyruvate dehydrogenase subunits PDHA1 and PDHA2. PDH phosphorylation inhibits pyruvate dehydrogenase activity, and thereby regulates metabolite flux through the tricarboxylic acid cycle, down-regulates aerobic respiration and inhibits the formation of acetyl-coenzyme A from pyruvate. Inhibition of pyruvate dehydrogenase decreases glucose utilization and increases fat metabolism in response to prolonged fasting and starvation. PDK4 plays an important role in maintaining normal blood glucose levels under starvation, and is involved in the insulin signaling cascade. Via its regulation of pyruvate dehydrogenase activity, PDK4 plays an important role in maintaining normal blood pH and in preventing the accumulation of ketone bodies under starvation. In the fed state, PDK4 mediates cellular responses to glucose levels and to a high-fat diet. PDK4 regulates both fatty acid oxidation and de novo fatty acid biosynthesis. PDK4 plays a role in the generation of reactive oxygen species.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab215544 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Interpolated concentrations of native PDK4 in C2C12 cell extract sample based on a 500 μg/mL extract load, mouse skeletal muscle tissue extract based on a 500 μg/mL extract load, and mouse and rat heart tissue extract based on a 300 μg/mL extract load. The concentrations of PDK4 were measured in duplicate and interpolated from the PDK4 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean PDK4 concentration was determined to be 4.8 ng/mL in C2C12 extract, 8.4 ng/mL in mouse skeletal muscle extract, 7.8 ng/mL in mouse heart extract, and 10.5 ng/mL in rat heart extract.

  • Interpolated concentrations of PDK4 in various cell and tissue extracts. The concentrations of PDK4 were measured in three different dilutions in duplicate and interpolated from the PDK4 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted in ng PDK4 per mg of extract (mean +/- SD, n=3). PDK4 concentration was determined to be 9.9 ng/mg in C2C12 cell extract, 17.3 ng/mg in mouse skeletal muscle extract, 26.7 ng/mg in mouse heart, and 36.7 ng/mg in rat heart.

  • Other species reactivity was determined by measuring a 300 µg/mL extract load of various species heart tissue extract, and interpolating the protein concentrations from the mouse standard curve.

Protocols

References

ab215544 has not yet been referenced specifically in any publications.

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