KDM6A/KDM6B Activity Quantification Assay Kit (Fluorometric) (ab156911)
Key features and details
- Assay type: Enzyme activity
- Detection method: Fluorescent
- Platform: Microplate reader
- Assay time: 3 hr
- Sample type: Nuclear Extracts, Purified protein
- Sensitivity: 10 ng
Overview
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Product name
KDM6A/KDM6B Activity Quantification Assay Kit (Fluorometric)
See all KDM6A/KDM6B kits -
Detection method
Fluorescent -
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.
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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.
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Platform
Microplate reader
Properties
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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 Assay Buffer 1 x 4ml 1 x 8ml Assay Standard, 50 µg/mL 1 x 10µl 1 x 20µl Substrate, 50 µg/mL 1 x 60µl 1 x 120µl -
Research areas
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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
see all
Images
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Typical Results
Demonstration of high sensitivity of KDM6A/KDM6B activity assay achieved by using KDM6A recombinant protein with the KDM6A/KDM6B Activity Quantification Assay Kit (Fluorometric).
