Application Note: Alpha Terbium SureFire Ultra Multiplex: simultaneous dual phosphoprotein target analysis

In this application note, PerkinElmer demonstrates the measurement of two phosphoprotein targets in a single sample using the  Alpha Terbium SureFire Ultra kit combined with the AlphaLISA SureFire Ultra kit…

Protein phosphorylation is a very useful readout of cellular activation or inhibition, and these pathways are commonly targeted for therapeutic modulation of disease. Examples include modulation of cell surface receptors, such as GPCRs or receptor tyrosine kinases, by antibodies or small molecules, and of intracellular kinases by cell permeable drugs.  Furthermore, measurement of more than one cellular target can provide greater confidence of positive compound hits on the cellular target of interest, through examination of an unrelated pathway or an upstream target that should be unaffected by the drug. The ability to measure more than one target within a single well of an assay in combination with the benefits of a homogenous, high throughput assay would be very valuable to screening programs.

The Alpha SureFire Ultra Multiplex assay was developed to do just that.  This assay is the only phosphoprotein multiplexing platform available that is truly homogenous and high throughput. The Alpha SureFire Ultra Multiplex assays are divided into two categories.  Here we describe the implementation of multiplexing using a combination of these two types of kits, the Terbium SureFire Ultra and the AlphaLISA SureFire Ultra kit, and provide example data to show assay performance.