Overview

  • Product name
    Human Transketolase ELISA Kit (TKT)
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    HeLa extract 5 2.3%
    Inter-assay
    Sample n Mean SD CV%
    HeLa extract 3 9.2%
  • Sample type
    Cell culture extracts, Tissue Extracts
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    800 pg/ml
  • Range
    4.68 ng/ml - 300 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 109.5 107.2% - 114%
    Cell culture media 114 111.9% - 117.7%
    Goat Serum 74.6 73% - 75.8%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
  • Product overview

    Transketolase (TKT) in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Transketolase protein in Human 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

    Transketolase is a key enzyme in the non-oxidative branch of the pentose phosphate pathway that transfers a two-carbon glycoaldehyde unit from ketose-donor to aldose-acceptor sugars. The enzyme is also involved in the photosynthetic Calvin cycle in plants and autotrophic bacteria. Thiamine diphosphate and calcium are essential cofactors in Transketolase catalyzed reactions. In mammals, Transketolase connects the pentose phosphate pathway to glycolysis, feeding excess sugar phosphates into the main carbohydrate metabolic pathways. Its presence is necessary for the production of NADPH, especially in tissues actively engaged in biosyntheses, such as fatty acid synthesis by the liver and mammary glands, and for steroid synthesis by the liver and adrenal glands. Transketolase activity is decreased in deficiency of thiamine, which is generally due to malnutrition, i.e. Wernicke-Korsakoff syndrome.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate

Properties

Applications

Our Abpromise guarantee covers the use of ab187398 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

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

  • Background-subtracted data values (mean +/- SD, n = 2) are graphed.

  • Background-subtracted data values (mean +/- SD, n = 2) are graphed.

  • Three concentrations (within the working range of the assay) of the cell extracts were analyzed in duplicates with this kit. The concentrations of TKT were interpolated from data values using TKT standard curve and graphed in ng of TKT per mg of extract (mean +/- SD, n=3).

Protocols

References

ab187398 has not yet been referenced specifically in any publications.

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