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AlphaLISA® Acceptor beads designed to detect human Histone H3 that is methylated at arginine 2(H3R2me) in a homogeneous AlphaLISA assay.
In the AlphaLISA assay, a biotinylated Histone H3 peptide substrate is used in a biochemical enzymatic reaction with histone methyltransferase (HMT) and S-adenosylmethionine (SAM) cofactor. When the enzymatic reaction is stopped, the level of methylation is determined using antibody-coated AlphaLISA Acceptor beads and streptavidin-coated Alpha Donor beads. The streptavidin Donor beads capture the biotin moeity on the peptide substrate, while the antibody on the AlphaLISA Acceptor beads recognizes the specific epigenetic mark (methylation of Histone H3 on arginine 2). Alternatively, a biotinylated Histone H3 (C-term) antibody (Cat. No. AL118) can be used to capture full-length Histone H3 substrates to the streptavidin Donor beads. Upon laser irradiation of the bead complexes at 680 nm, short-lived singlet oxygen molecules produced by the Donor beads can reach the Acceptor beads in proximity to generate an amplified chemiluminescent signal at 615 nm. The intensity of light emission is proportional to the level of substrate modification.
|Bead Type or Core Bead Type||AlphaLISA Acceptor|
|Experimental Type||In vitro|
|Product Brand Name||AlphaLISA|
|Shipping Condition||Blue Ice|
|Unit Size||250 µg|
AlphaLISA technology is a powerful and versatile platform that offers, highly sensitive, no-wash immunoassays using Alpha Donor and, AlphaLISA Acceptor beads. In this technical note, we present the, optimization of a PRMT6 enzymatic assay using a biotinylated histone, H3-derived peptide as substrate. Detection of the methylated product, was performed by the addition of Streptavidin (SA) Alpha Donor, beads and AlphaLISA Acceptor beads conjugated to an antibody (Ab), directed against the methylated H3R2 residue. Upon laser irradiation, of the beads-target complexes at 680 nm, short-lived singlet oxygen, molecules produced by the Donor beads can reach the Acceptor, beads in proximity to generate an amplified chemiluminescent signal, at 615 nm. The intensity of the light emission is proportional to the, methylation activity of the PRMT6 enzyme.