Overview

  • Product name

    Human Hexokinase II ELISA Kit
    See all Hexokinase II kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    HeLa extract 8 3%
    Inter-assay
    Sample n Mean SD CV%
    HeLa extract 3 5.7%
  • Sample type

    Serum, Cell culture extracts, Tissue Extracts, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    11.37 pg/ml
  • Range

    62.5 pg/ml - 4000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 99 96% - 102%
    Cell culture extracts 92 89% - 102%
    Tissue Extracts 96 91% - 103%
    Heparin Plasma 101 98% - 106%
    EDTA Plasma 102 97% - 106%
    Citrate Plasma 96 91% - 99%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Product overview

    Hexokinase II in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Hexokinase II protein in serum, plasma, and 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.


    Sensitivity: 


    Samples in Sample Diluent 10BS – 17.58 pg/mL
    Samples in 1X Cell Extraction Buffer PTR – 11.37 pg/mL

  • Notes

    Hexokinases are involved in the pathway of hexose metabolism, where they phosphorylate hexose to form hexose-6-phosphate. Phosphorylation of hexose restricts transport out of the cell, therefore retained intracellularly for metabolic processes including glycolysis and glycogen synthesis. Hexokinases also phosphorylate glucose to glucose-6-phosphate.

    There are four mammalian hexokinases. Hexokinase II, also known as muscle form hexokinase, localizes to the outer membrane of mitochondria and is present in adipose tissue, skeletal muscle, and heart. Hexokinase II comprises of two homologous halves with both the C-terminal and N-terminal portions being catalytically active (with the N-terminal half having higher activity than the C-terminal half). Human Hexokinase I, mouse Hexokinase II, and rat Hexokinase II share 73.8%, 94.2%, and 94.4% in homology in amino acid sequence with human Hexokinase II, respectively.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

Applications

Our Abpromise guarantee covers the use of ab219043 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.

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

  • The concentrations of Hexokinase II were measured in duplicates, interpolated from the Hexokinase II standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (citrate) 50%, plasma (EDTA) 50%, and plasma (heparin) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Hexokinase II concentration was determined to be 7,009.26 pg/mL in neat serum, 7,887.82 pg/mL in neat plasma (citrate), 8,052.65 pg/mL in neat plasma (EDTA), and 8,675.63 pg/mL in neat plasma (heparin).

  • The concentrations of Hexokinase II were measured in duplicate and interpolated from the Hexokinase II standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Hexokinase II concentration was determined to be 1,175.25 pg/mL in placenta extract, 2,844.72 pg/mL in HeLa extract, and 2,577.06 pg/mL in Jurkat extract.

     

     

  • The concentrations of Hexokinase II were measured in duplicate and interpolated from the Hexokinase II standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Hexokinase II concentration was determined to be 624.25 pg/mL in human fetal brain extract, 4,691.00 pg/mL in mouse brain extract, and 973.64 pg/mL in rat brain extract.

Protocols

References

ab219043 has not yet been referenced specifically in any publications.

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