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The LANCE® Ultra Europium-anti-unmodified Histone H3 Lysine 9/Lysine 27 (H3K9/K27) antibody was used for the development and optimization of epigenetic writer and eraser 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||Dry Ice|
|Unit Size||100 µg|
The introduction of enzyme-linked immunosorbent assays (ELISAs) in the early 1970’s offered researchers a non-radiometric immunoassay platform without compromising sensitivity. Over the last 50 years scientists have made huge strides in disease research and drug discovery and a demand for greater assay throughput and sensitivity has evolved. In response, more robust immunoassays have been developed to address some of the limitations of the standard, colorimetric ELISA.
Find out about the most common limitations of traditional ELISAs and how different ELISA alternative technologies address these limitations.
In this technical note, we present the optimization of an HDAC1, enzymatic assay using as substrate a biotinylated histone H3-derived, peptide acetylated at lysine 9. The deacetylated peptide product is, captured by the Eu-labeled antibody (Eu-Ab) and ULight-Streptavidin, (ULight-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 deacetylation activity of the, HDAC1 enzyme.
LANCE Ultra time-resolved fluorescence resonance energy transfer, (TR-FRET) assays use a proprietary europium chelate donor dye, W1024 (Eu), together with ULight™, a small molecular weight, acceptor dye with a red-shifted fluorescent emission.