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Simultaneous Detection of Total and Phosphorylated Protein in Cellular Kinase Assays using Alpha SureFire Ultra Multiplex Technology

PerkinElmer
31 Jul 2017

Cellular kinase signal transduction pathways are involved in the regulation of many important cellular processes such as, cell survival, differentiation, and apoptosis. Numerous assay technologies have been developed for studying kinase signaling pathways, and for screening compound libraries in search of agents to modify receptors or kinase activities. Alpha SureFire® Ultra™ Multiplex Phospho and Total assays utilize two types of Alpha Acceptor beads to simultaneously measure two signals from each assay well to easily normalize the assay. This application note demonstrates the benefit and utility of normalizing the assay signal of phosphorylated protein levels to total protein levels in two different cellular models.

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Alpha is a bead-based, no-wash luminescent technology. When Alpha Donor and Acceptor beads are brought together through binding to target, a cascade of chemical reactions is set in motion, creating an amplified signal. These versatile beads are conjugated with various biomolecules that enable detection of unique biological events, offering maximum flexibility.

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