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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.
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.
|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|
Our Abpromise guarantee covers the use of ab156911 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Functional Studies||Use at an assay dependent concentration.|
ab156911 has not yet been referenced specifically in any publications.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"