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The LANCE® Ultra Europium-anti-tri-methyl-Histone H3 Lysine 27 (H3K27me3) antibody was used for the development and optimization of methyltransferase assays.
Proven, cost-efficient LANCE Ultra reagents can be used to quantitate peptide modifications, detecting specific meythylation and acetylation states. No wash, homogenous LANCE Ultra reagents are HTS friendly - design your own epigenetics screening strategy for greatest efficiency.
Epigenetic enzymatic assays are optimized using a biotinylated histone H3-derived peptide as substrate. The modified peptide is captured by the Eu-labeled antibody (Ab) and ULight-Streptavidin (SA) which bring the Eu donor and ULight acceptor dye molecules into close proximity. Upon irradiation at 320 or 340 nm, the energy from the Eu donor is transferred to the ULight acceptor dye which, in turn, generates light at 665 nm. The intensity of the light emission is proportional to the level of biotinylated substrate modification.
|Detection Method||Time-Resolved Fluorescence (TRF), TR-FRET|
|Experimental Type||In vitro|
|Product Brand Name||LANCE Ultra|
|Shipping Condition||Blue Ice|
|Unit Size||100 µg|
Anti-mark antibodies coupled to AlphaLISA Acceptor beads or labeled with LANCE Ultra europium chelate were used for the successful optimization of robust and, sensitive epigenetic assays using histone H3-derived peptides as substrates.
In eukaryotes, the covalent modification of histones has a crucial role in chromatin architecture and plays an important part in a plethora of cellular processes, from chromatinre modeling and transcriptional regulation, to DNA repair and cell cycle control.
Covalen modification of DNA through methylation is catalyzed by specific DNA methyltransferases (DNMTs).
In this technical note, we present the optimization of an epigenetic enzymatic assay using a biotinylated histone H3-derived peptide as substrate. The modified peptide is captured by the Eu-labeled antibody(Eu-Ab) and ULight-Streptavidin (SA) which bring the Eu donor and ULight acceptor dye molecules into close proximity.