Histone Methylation with Activity Assays

Measure histone methyltransferase and demethylase activity with a variety of solutions offered by Abcam

Histone Methyltransferase Activity Assays

The development of assays for histone methyltransferases (HMTs) is hard because they have often had low kcat, with a turnover that is usually below 1 min-1, and low KM values for their methyl donor S-adenosylmethionine (SAM). It is mandatory, therefore, for HMT assays to have very high sensitivity.

Many HMT activity assays measure S-adenosylhomocysteine (SAH) formation, a by-product of the methylation reaction. This makes them unspecific for any HMT and it is only possible to use them with purified protein/protein complexes. They also often use 3H-SAM as a donor, and so involve handling radioactive material.

Abcam’s HMT activity assays handle those challenges by using antibodies to detect the methylated product, providing:

  • Quantitative detection that is easy and does not need radioactivity
  • As the assay is specific for the modification of interest, the ability to use nuclear extracts as well as purified proteins is applied
  • Microplate format with simple colorimetric or fluorometric readouts
  • Data in 3 hours ​​​​
Activity measured Product name Product code
Methylation of histone
H3 on K4
Histone H3 (K4) Methyltransferase Activity Quantification Assay Kit ab113452  
Methylation of histone
H3 on K9
Histone H3 (K9) Methyltransferase Activity Quantification Assay Kit ab113453  
Methylation of histone
H3 on K27
Histone H3 (K27) Methyltransferase Activity Quantification Assay Kit ab113454  

Histone Demethylase Activity Assays

The formation of formaldehyde is typically measured by histone demethylase assays, a by-product of demethylation. They can therefore be easily interfered with by detergents, thiol groups and a range of ions. Furthermore, these assays are not specific for any demethylase and can only be run with purified protein.

The histone demethylase assays measure directly the formation of demethylated product, thus avoiding the issues, and providing:

  • Increased sensitivity (20–1000 fold) over formaldehyde-based assays
  • More precise data – no interference from thiols, detergents or ions
  • Compatibility with nuclear extracts as well as purified protein (due to the assay’s specificity for the modification of interest)
  • Microplate format with simple colorimetric or fluorometric readouts
  • 3 hour assay time
Activity measured Product name Product code
Demethylation of histone H3K4me2 Histone Demethylase (H3K4)
Activity Quantification Assay Kit
ab113455
Demethylation of histone H3K4me2 and H3K4me3 by KDM1 / LSD1

KDM1/LSD1 Activity Quantification Assay Kit (Colorimetric)

KDM1/LSD1 Activity Quantification Assay Kit (Fluorometric)

ab113459

​                                          

ab113460

Demethylation of H3K9me3 by: KDM4A/JHDM3A/JMJD2A, KDM4B/JMJD2B, KDM4C/GASC1/JMJD2C and KMD4D/JMJD2D

KDM4/JMJD2 Activity Quantification Assay Kit (Colorimetric)

KDM4/JMJD2 Activity Quantification Assay Kit (Fluorometric)

ab113461

ab113462

Demethylation of H3K4me2 and H3K4me3 by KDM5/JARID

KDM5/JARID Activity Quantification Assay Kit (Colorimetric)

KDM5/JARID Activity Quantification Assay Kit (Fluorometric)

ab113463       ​

ab113464

Demethylation of H3K27me3 by KDM6A/
UTX and KDM6B/JMJD3

KDM6A/KDM6B Activity Quantification Assay Kit (Colorimetric)

KDM6A/KDM6B Activity Quantification Assay Kit (Fluorometric)

ab156910

     

ab156911

About Abcam

Abcam is a global life sciences company providing highly validated antibodies and other binders and assays to the research and clinical communities to help advance the understanding of biology and causes of disease.

Abcam’s mission is to serve life scientists to help them achieve their mission faster by listening to their needs, continuously innovating and improving and by giving them the tools, data and experience they want. Abcam’s ambition is to become the most influential life science company for researchers worldwide.


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Last updated: May 24, 2019 at 8:01 AM

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