AlphaScreen phosphoprotein kits are designed to detection phosphoproteins in biochemical or cellular kinase assays. This kit contains streptavidin Donor beads and AlphaScreen Acceptor beads conjugated with the antiphosphotyrosine P-Tyr-100 antibody. This kit can be used to capture biotinylated phosphoproteins, biotinylated phosphorylated peptides, or biotinylated antibodies in complex with phosphorylated proteins.
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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.
|Product Brand Name||AlphaScreen|
|Shipping Condition||Blue Ice|
|Unit Size||500 assay points|
Protein tyrosine kinases play a key role in signal transduction and normal cell growth. They are also involved in numerous proliferative diseases like cancer and atherosclerosis, in addition to anumber of autoimmune diseases. For that reason the tyrosine kinases are often targeted in researchand HTS laboratories for new drug discovery.