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These beads can be used to capture GST-tagged proteins and biomolecules, and can be used in conjunction with AlphaScreen, AlphaLISA, or AlphaPlex Acceptor beads to create no-wash assays for:
In a typical Alpha assay, 1 mg of Donor beads is sufficient to run 1,000-2,000 wells using a 25 µL reaction volume. Bead concentration can be adjusted for optimal performance.
|Bead Type or Core Bead Type||Alpha Donor|
|Product Brand Name||AlphaLISA|
|Shipping Condition||Blue Ice|
|Unit Size||1 mg|
Product brochure for the Alpha Technology, a versatile, no wash, homogeneous assay technology that's suitable for a broad range of applications.
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.
This guide presents the simple conversion of an ELISA or other immunoassay to an AlphaLISA® immunoassay.
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. The assay does not require any washing steps. Binding of proteins or other binding partners captured on the beads leads to an energy transfer from one bead to the other, ultimately producing a luminescent signal.
Mitogen-activated protein kinases, MAPKs, are constituents of numerous signal transduction pathways, and are activated by protein kinase cascades. There is great interest in finding compounds that target components of the MAPK pathways as possible treatment for inflammatory diseases and cancer. Here we report the development of a novel AlphaScreen® assay to measure MEK2 mediated phosphorylation of Erk-1.
Protein kinases are directly implicated in many human diseases, including inflammatory diseases and cancer; thus, kinases are promising therapeutic targets. In kinase screening and deorphanization studies, the use of protein substrates constitutes a major advantage over peptide substrates. The AlphaScreen® PhosphoSensor™ GST kit was developed to simplify kinase assays using physiological substrates.
Binding events between biomolecules are important components of biological processes and a number of these biomolecular interactions have been targeted for the development of novel therapeutic drugs. p53 is a transcription factor and tumor suppressor protein that is activated in response to cellular stress, and MDM2 was identified as a negative regulator that binds to p53 and tags it for ubiquitination and subsequent degradation.
The p53-MDM2 protein-protein interaction has been an excellent target for therapeutic drugs and therefore makes a good model system for developing an AlphaLISA assay to screen for inhibitors of the interaction. In this technical note, we show how to develop an assay to screen for inhibitors and how to measure a dissociation constant for moderate binding protein-protein interaction using AlphaLISA®.