EPIgeneous Methyltransferase kit HTRF®
The EPIgeneous methyltransferase assay is a kit optimized for the assessment of enzymatic methylation activity through S-adenosylhomocysteine detection (SAH).
- High sensitivity
The EPIgeneous Methyltransferase assay is a universal solution for all methyltransferases and substrates (peptides, nucleosomes, full histones, DNA, RNA, p53 and small molecules). The detection is made via the measurement s-adenosylhomocysteine (SAH) from s-adenosylmethinionine (SAM) conversion by methyltransferases. The assay is compatible with most enzyme with a Km for SAM between 0.4 µM 200 µM. The assay provides a unique and robust alternative to radioactivity.
- HTS AND PROFILING
- ROBUST & READY TO USE
Enzymatic step principle
Detection step principle
DOT1L Enzyme titration
Human recombinant DOT1L was serially diluted in enzymatic buffer from 320 nM to 0.001 nM. The assay carried out with 10 ng/µl (= 77 nM) oligonucleosome as substrate and 2 µM SAM for 2 h at 30°C. The negative controls (no SAM or no nucleosome) confirm the measurement the enzymatic specific activity. A DOT1L concentration of 4.5 nM (EC80) was selected for further experiments. This concentration leads to 10% conversion of SAM into SAH.
Inhibition effect of SGC0946 on DOT1L assay
The graph represents the assay windows obtained with a range of SAM concentrations and at different conversion percentages of SAM into SAH. The assay was performed using the calibration curve of the kit prepared with the assay diluent The EPIgeneous methyltransferase assay has been optimized to be suitable for a large range of SAM concentrations in the enzymatic step (0.4 – 200 µM). With these concentrations of SAM, the assay is able to assess the enzymatic activity with 4 to 10% turnover of the enzyme (hence 4 to 10% conversion of SAM into SAH).
Automation of a Generic Fluorescence Methyltransferase Activity Assay
In collaboration with BioTek - Posters
HTRF Universal SAH detection assay to assess methyltransferase activity
DOT1L/oligonucleosome assay - Posters
Setting-up HTRF technology for G9a & DNMT1 High-Throughput Screening
In collaboration with Universidad de Navarra - Posters
How HTRF compares to Western Blot and ELISA
Get the brochure about technology comparison. - Brochures
Plate Reader Requirement
Choosing the right microplate reader ensures you’ll get an optimal readout. Discover our high performance reader, or verify if your lab equipment is going to be compatible with this detection technology.Let's find your reader