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  • Application Note

    Measuring PD-L1 Expression in Breast Cancer Cell Lines with AlphaLISA

    Too many candidates, too little time. The lack of robust, rapid, high-throughput assays to identify and qualify potential therapeutic targets in areas such as cancer research continues to cost valuable time. What if you could increase assay throughput without compromising sensitivity, obtain more data points from each sample and eliminate tedious wash steps? Find out how AlphaLISA® assay technology, combined with the EnVision® multimode plate reader, provides a fast, powerful, homogeneous platform for screening potential inhibitors of PD-L1 (a protein associated with breast cancer tumor cells) expression in human cells.

  • Application Note

    Simultaneous Detection of Total and Phosphorylated Protein in Cellular Kinase Assay Using Alpha SureFire Ultra Multiplex Technology

    Cellular kinase signal transduction pathways are involved in the regulation of many important cellular processes such as cell survival, differentiation, and apoptosis. Kinase signaling networks are typically characterized by multiple kinases arranged in cascades containing nodes with feedback loops and crosstalk between pathways. 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. The quality of these assays can be impacted by data variability due to cell seeding inhomogeneity or from compound toxicity. A common method for controlling for variability is to normalize the assay signal to a cellular protein whose level does not change as a function of the treatment.

    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. Here, we demonstrate the benefit and utility of normalizing assay signal of phosphorylated protein levels to total protein levels in two different cellular models: ERK 1/2 phosphorylation in human melanoma A375 cells and AKT 1/2/3 phosphorylation in mouse myoblast C2C12 cells.