Key features and details
- Assay type: Enzyme activity
- Detection method: Fluorescent
- Platform: Microplate reader
- Sample type: Purified protein
Product nameUniversal Methyltransferase Activity Assay Kit
See all Methyltransferase kits
Sample typePurified protein
Assay typeEnzyme activity
Universal Methyltransferase Activity Assay Kit (ab139433) is a homogeneous mix-and-read fluorescent assay for the determination of any S-adenosyl-L-methionine (AdoMet/SAM) dependent methyltransferase activity. It is suitable for end-point or kinetic read options, which is ideal for determining mechanism of action, kinetics, and screening candidate compounds. The assay is amendable to HTS and miniaturization.
ab139433 is a complete kit for the screening of candidate compounds that may alter normal methyltransferase activity.
Methylation of proteins, nucleic acids and oligosaccharides is an important post- translational regulatory event. Activities that are methylation-related include meiosis, biosynthesis, development, signal transduction, chromatin regulation, and gene silencing. The enzymes that mediate the covalent transfer of a methyl group from a donor to an acceptor molecule are methyltransferases. Methyltransferases have structurally unrelated acceptors as diverse as proteins and DNA, however frequently use S -adenosylmethionine as a universal donor. Part of the acceptor diversity of this enzyme family relates to the flexible structural folds that bind these molecules in proximity of the donor. The side- chains of lysine, arginine, glutamate, glutamine, asparagines, and isoprenylated residues serve as methylation sites in proteins like histones. Changes in methylation patterns have been tightly linked to disease states such as cancer and vascular disease.
Storage instructionsPlease refer to protocols.
Components 1 x 96 tests Black Microtiter Plate 1 unit Methyltransferase Positive Control 1 x 0.2ml Methyltransferase Reaction Buffer Concentrate 1 x 0.3ml Plate Sealer 1 x 3 units Transferase Assay Buffer Concentrate 1 x 15ml Transferase Detection Solution Concentrate 1 x 0.15ml
Using SET7/9 and TAF-10 as an enzyme substrate system, percent inhibition for dilutions of 5’deoxy-5’(methylthio)-adenosine was tested.
0.5µM SET7/9 was tested with 15µM TAF-10 substrate in the kinetic assay format. Mean relative fluorescence was plotted against the stop time interval.
Titration assay of an engineered portion of human lysine specific histone methyltransferase SET7/9 using 15µM of the peptide substrate TAF-10. Serial dilutions of the enzyme were prepared in Transferase Assay Buffer. Average relative fluorescence was plotted against SET7/9 concentration to generate the following graph. This is for maximum signal within the detection range of the plate reader, with a signal to noise ratio sufficient for easy detection of altered enzyme activity.
ab139433 has not yet been referenced specifically in any publications.