Corporate Banner
Satellite Banner
Scientific Community
Become a Member | Sign in
Home>Resources>Application Notes>This Application Note
  Application Notes
Scientific News
Researchers Develop Classification Model for Cancers Caused by KRAS
Most frequently mutated cancer gene help oncologists choose more effective cancer therapies.
Chromosomal Chaos
Penn study forms basis for future precision medicine approaches for Sezary syndrome
Shaking Up the Foundations of Epigenetics
Researchers at the Centre for Genomic Regulation (CRG) and the University of Barcelona (UB) published a study that challenges some of the current beliefs about epigenetics.
Genetic Defences of Bacteria Don’t Aid Antibiotic Resistance
Genetic responses to the stresses caused by antibiotics don’t help bacteria to evolve a resistance to the medications, according to a new study by Oxford University researchers.
Tolerant Immune System Increases Cancer Risk
Researchers have found that individuals with high immunoCRIT ratios may have an increased risk of developing certain cancers.
Developing a Gel that Mimics Human Breast for Cancer Research
Scientists at the Universities of Manchester and Nottingham have been funded to develop a gel that will match many of the biological structures of human breast tissue, to advance cancer research and reduce animal testing.
Lung Repair and Regeneration Gene Discovered
New role for hedgehog gene offers better understanding of lung disease.
3 Ways Viruses Have Changed Science for the Better
Viruses are really good at what they do, and we’ve been able to harness their skills to learn about – and potentially improve – human health in several ways.
Mixed Up Cell Transportation Key Piece of ALS and Dementia Puzzle
Researchers from the University of Toronto are one step closer to solving this incredibly complex puzzle, offering hope for treatment.
New Gene Therapy for Vision Loss From a Mitochondrial Disease
NIH-funded study shows success in targeting mitochondrial DNA in mice.
Scroll Up
Scroll Down

Automation of a Generic Fluorescent Methyltransferase Activity Assay
Bookmark and Share

BioTek Instruments


Epigenetic processes are attracting considerable attention in drug discovery as their fundamental roles in controlling normal cell development and contributions to disease states become more clearly defined. Methylation is a known ubiquitous covalent modification involved in regulation of a diverse range of biomolecules. As histone methylation is linked to certain disease states, including many cancer types, Histone methyltransferases (HMTs) are of particular interest as drug targets. A high-throughput screening (HTS)-ready, universal methyltransferase activity assay was recently developed based on competitive fluorescent polarization immunodetection of adenosine monophosphate (AMP), formed from the methyltransferase (MT) reaction product S-adenosylhomocysteine (SAH) in a dual enzyme coupling step.

Here we demonstrate automation of the assay in a 384-well microplate format suitable for HTS. A combination 8-channel and single channel liquid handling instrument was used to automate histone methyltransferase (HMT) G9a titration and transfer to the assay plate in quadruplicate. Automated serial dilution and transfer of the HMT inhibitor sinefungin was also performed using the liquid handler’s 8-channel head. During HMT EC80 determination, cofactor, substrate, stop buffer and detection reagent additions were automated using a non-contact dispenser. Dispensing of the EC80 concentration of HMT enzyme and all assay components were automated during HMT inhibitor IC50 determination. The G9a EC80 was determined to be approximately 80 ng/mL and used for subsequent inhibition studies, while the sinefungin IC50 value was determined to be 14.5 μM, showing excellent correlation with published values.

Further Information

Related Content

DLReady Certification Awarded for Synergy 2 and Synergy HT Multi-Detection Microplate Readers
Awarded by Promega Corporation, this certification validates an instrument to the highest performance levels and standards for the Dual-Luciferase® Reporter Assay System.
Monday, November 06, 2006
Skyscraper Banner

Skyscraper Banner
Go to LabTube
Go to eposters
Access to the latest scientific news
Exclusive articles
Upload and share your posters on ePosters
Latest presentations and webinars
View a library of 1,800+ scientific and medical posters
2,600+ scientific and medical posters
A library of 2,500+ scientific videos on LabTube
3,800+ scientific videos