KDM6A/KDM6B Activity Quantification Assay Kit (Fluorometric) (ab156911)

Overview

  • Product name
    KDM6A/KDM6B Activity Quantification Assay Kit (Fluorometric)
    See all KDM6A/KDM6B kits
  • Sample type
    Nuclear Extracts, Purified protein
  • Assay type
    Enzyme activity
  • Sensitivity
    10 ng
  • Assay time
    3h 00m
  • Species reactivity
    Reacts with: Plants, Mammals, Fungi
  • Product overview

    KDM6A/KDM6B Activity Quantification Assay Kit (Fluorometric) (ab156911) is a complete set of optimized reagents, designed for measuring activity/inhibition of KDM6A and KDM6B using nuclear extracts or purified enzymes from a broad range of species such as mammals, plants, fungi, and bacteria, in a variety of forms including, but not limited to cultured cells and fresh and frozen tissues. There are currently a limited number of methods used for detecting KDM6A/KDM6B activity/inhibition.

    The traditional method is based on the measurement of formaldehyde release, a by-product of KDM6A/KDM6B enzymatic reaction, and has significant weaknesses: (1) large amounts (at µg level) of substrate and enzyme are required; (2) nuclear extracts from cell/tissues cannot be used; (3) redox-sensitive KDM6A/KDM6B inhibitiors are not suitable for testing with such methods; (4) high intereference by SDS, DMSO, thiol-containing chemicals, and ions, which are often contained in enzyme solutions, tested compound solvents, and assay buffers; and (5) less accurate than direct measurement of KDM6A/KDM6B-converted demethylated products.

  • Notes

    Lysine specific demethylase 6A (KDM6A/UTX) and lysine specific demethylase 6B (KDM6B/JMJD3) demethylase lysine 27 on histone H3, thereby playing a central role in histone code. KDM6A and KDM6B are JmjC (Jumonji)-domain-containing proteins and catalyze the removal of di- and tri-methylation from histone H3-K27 by using a hydroxylation reaction with iron and alpha-glutarate as cofactors.

Properties

  • Storage instructions
    Please refer to protocols.
  • Components 48 tests 96 tests
    10X Wash Buffer 1 x 14ml 1 x 28ml
    8-Well Assay Strips (with Frame) 1 x 6 units 1 x 12 units
    Adhesive Covering Film 1 unit 1 unit
    Capture Antibody, 1000 µg/mL 1 x 5µl 1 x 10µl
    Co-factor 1 1 x 30µl 1 x 60µl
    Co-factor 2 1 x 30µl 1 x 60µl
    Co-factor 3 1 x 30µl 1 x 60µl
    Detection Antibody, 400 µg/mL 1 x 6µl 1 x 12µl
    Dilution Buffer 1 x 4ml 1 x 8ml
    Fluoro-Developer 1 x 10µl 1 x 20µl
    Fluoro-Enhancer 1 x 10µl 1 x 20µl
    JMJD Assay Buffer 1 x 4ml 1 x 8ml
    JMJD Assay Standard, 50 µg/mL 1 x 10µl 1 x 20µl
    JMJD Substrate, 50 µg/mL 1 x 60µl 1 x 120µl
  • Research areas
  • Relevance
    Histone demethylase that specifically demethylates 'Lys-27' of histone H3, thereby playing a central role in histone code. Demethylates trimethylated and dimethylated H3 'Lys-27'. Plays a central role in regulation of posterior development, by regulating HOX gene expression.
  • Cellular localization
    Nuclear
  • Alternative names
    • bA386N14.2
    • Histone demethylase UTX
    • JmjC domain containing protein 3
    • JMJD3
    • Jumonji domain containing 3, histone lysine demethylase
    • Jumonji domain containing protein 3
    • KABUK2
    • KDM6A
    • KDM6B
    • Lysine (K) specific demethylase 6A
    • Lysine (K) specific demethylase 6B
    • Lysine demethylase 6B
    • Lysine specific demethylase 6A
    • Lysine specific demethylase 6B
    • Ubiquitously transcribed tetratricopeptide repeat protein X linked
    • Ubiquitously transcribed TPR gene on the X chromosome
    • Ubiquitously transcribed TPR protein on the X chromosome
    • Ubiquitously transcribed X chromosome tetratricopeptide repeat protein
    • UTX
    see all

Associated products

Images

  • Demonstration of high sensitivity of KDM6A/KDM6B activity assay achieved by using KDM6A recombinant protein with the  KDM6A/KDM6B Activity Quantification Assay Kit (Fluorometric).

Protocols

References

ab156911 has not yet been referenced specifically in any publications.

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