Revvity, 62SAHPEB, HTRF EPIgeneous Methyltransferase Assay Kit, 1,000 Assay Points

Overview
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.
HTRF assays offer many advantages over other technologies:

Homogeneous add-and-read format
No wash steps
Low background
Straightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates
Stable signal, providing flexibility in time of readout or size of assays
How it works
Enzymatic step principle
The EPIgeneous Methyltransferase Assay is a universal biochemical assay for all enzymes within the histone (HMTs) and DNA (DNMTs) methyltransferase families that produce S-adenosylhomocysteine (SAH). The methyltransferase activity is assessed by measuring the conversion of SAM (S-5adenosyl-L-methionine) to SAH.

 

Detection step principle
In order to directly measure SAH release, an anti SAH antibody labeled with Tb cryptate and a SAH-d2 tracer are used. The SAH released by the enzymatic reaction competes with the SAH-d2 labeled leading a decrease of the HTRF signal.

 

Assay protocol
The substrate is incubated with the enzyme in presence of compounds. SAM (CH3 donor) is added to start the enzymatic reaction leading to the substrate methylation. The antibody specific to SAH labeled with Tb cryptate competes with both native SAH and d2-coupled SAH. The resulting TR-FRET signal is inversely proportional to the concentration of SAH in the calibrator or in the sample. Application-Biochemical Enzymatic Assay
Brand-EPIgeneous
Detection Modality-HTRF
Product Group-Kit
Sample Volume-120 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Epigenetics
Technology-TR-FRET
Therapeutic Area-Metabolism/Diabetes Neuroscience Oncology & Inflammation
Unit Size-1,000 assay points