In vitro histone modification (HM) assays are used to characterize the activity of chromatin-modifying enzymes. These assays provide information regarding the modification sites on histones, the product specificity, and the impact of other histone or nucleotide modifications on enzyme activity.

This protocol describes an in vitro assay for histone methyltransferase activity that uses bacterial cell extracts in which expression of a methyltransferase of interest is induced. In many cases, purification of the enzyme is unnecessary, making this experiment ideal for pilot studies.

Feb 8, 2021 · Assay development for histone methyltransferases (HMTs) is difficult because they have often have low kcat (turnover is typically below 1 min-1) and low KM values for their methyl donor S-adenosylmethionine (SAM). HMT assays therefore have need to …

assays provide highly sensitive and robust options for universal detection of HMT activity and are the preferred option for profiling against panels of methyltransferases. Optimizing the radioisotope filter binding platform for HTS, Reaction

Jun 23, 2022 · In vitro histone methyltransferase assays have been instrumental in identifying HMTs, and they continue to be invaluable tools for the study of these important enzymes, revealing novel substrates and modes of regulation.

The EpiQuik™ Histone Methyltransferase Activity/Inhibition Assay Kit (H3-K27) is designed for measuring HMTs that specifically target histone H3 at lysine 27. In an assay with this kit, the histone substrate is stably captured on the strip wells through biotin-streptavidin binding.

In this study, we describe the application of a miniaturized, radioisotope-based reaction system: the HotSpot SM platform for methyltransferases. Since this platform employs tritiated SAM as a cofactor, it can be applied to the assay of any HMT.

In this study, we describe the application of a miniaturized, radioisotope-based reaction system: the HotSpot(SM) platform for methyltransferases. Since this platform employs tritiated SAM as a cofactor, it can be applied to the assay of any HMT.

(HTS) assay format, and reference inhibitors are not available for many of the enzymes. In this study, we describe the application of a minia­ turized, radioisotope-based reaction system: the HotSpofM platform for methyltransferases. Since this platform employs tritiated SAM as a cofactor, it can be applied to the assay of any HMT. The key ...

Histone methyltransferases (HMTs) catalyze the methylation of lysine and arginine residues in histone as well as nonhistone substrates. In vitro histone methyltransferase assays have been instrumental in identifying HMTs, and they continue to be invaluable tools for the study of these important enzy …