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AlphaLISA® Acceptor beads designed to detect human Histone H3 that is mono- and di-methylated at lysine 27 (H3K27me2-1) in a homogeneous AlphaLISA assay. Broad species cross-reactivity is expected based on sequence similarity. Source of antibody: monoclonal.
The anti-methyl-Histone H3 Lysine 27 (H3K27me2-1) AlphaLISA Acceptor beads were used for the development and optimization of an EZH2 methyltransferase assay using a biotinylated Histone H3 (21-44) as substrate. A JMJD3 demethylase assay was also developed, using a biotinylated Histone H3 (21-44) peptide trimethylated at lysine 27 as substrate. Technical notes describing both assays are available in our product literature.
AlphaScreen® and AlphaLISA® are bead-based assay technologies used to study biomolecular interactions in a microplate format. The acronym "Alpha" stands for amplified luminescent proximity homogeneous assay. As the name implies, some of the key features of these technologies are that they are non-radioactive, homogeneous proximity assays. Binding of molecules captured on the beads leads to an energy transfer from one bead to the other, ultimately producing a luminescent/fluorescent signal. To understand how a signal is produced, one must begin with an understanding of the beads. AlphaScreen and AlphaLISA assays require two bead types: Donor beads and Acceptor beads. Each bead type contains a different proprietary mixture of chemicals, which are key elements of the AlphaScreen technology. Donor beads contain a photosensitizer, phthalocyanine, which converts ambient oxygen to an excited and reactive form of O2, singlet oxygen, upon illumination at 680 nm. Please note that singlet oxygen is not a radical; it is molecular oxygen with a single excited electron. Like other excited molecules, singlet oxygen has a limited lifetime prior to falling back to ground state. Within its 4 µsec half-life, singlet oxygen can diffuse approximately 200 nm in solution. If an Acceptor bead is within that proximity, energy is transferred from the singlet oxygen to thioxene derivatives within the Acceptor bead, subsequently culminating in light production at 520-620 nm (AlphaScreen) or at 615 nm (AlphaLISA). In the absence of an Acceptor bead, singlet oxygen falls to ground state and no signal is produced. This proximity-dependent chemical energy transfer is the basis for AlphaScreen's homogeneous nature.
|Bead Type or Core Bead Type||AlphaLISA Acceptor|
|Experimental Type||In vitro|
|Product Brand Name||AlphaLISA|
|Shipping Condition||Blue Ice|
|Unit Size||250 µg|
The interactions and bindingof proteins are implicated in a large number of biological processes. The needfor an efficient, highly sensitive assay to study large protein interactions is increasingly important. Alpha Technology is a highly flexible, homogeneous, no-wash assay ideal for the measurement of protein interactions and complexes as large as 200 nm in size
Alpha has been used to study a wide variety of interactions, including protein:protein, protein:peptide, protein:DNA, protein:RNA, protein:carbohydrate, protein:small molecule, receptor:ligand, and nuclear receptor:ligand interactions. Both cell-based and biochemical interactions have been monitored, and applications such as phage display, ELISA, and EMSA (electrophoretic mobility shift assay) have been adapted to Alpha.
The AlphaLISA® assay is a homogeneous immunoassay alternative to classical ELISA. AlphaLISA assays were originally utilized to detect analytesin cell cultures upernatants or serum/plasma samples.
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).
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 an epigenetic enzymatic assay using a biotinylated Histone H3-derived peptide as substrate.
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 an epigenetic enzymatic assay using a biotinylated Histone H3-derived peptide as substrate.
The AlphaLISA technology allows performing no-wash homogeneous proximity immunoassays using Alpha Donor and AlphaLISA Acceptor beads. In this technical note, we present the optimization of anepigenetic enzymatic assay using a biotinylated histone H3-derived peptide as substrate.