SLAS

W526:Huwiler:MonitoringPostTranslationalModificationsAndProteinProteinInteractionsUsingTheProximityLigationAssay

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Kristin G. Huwiler, Tina Settineri, Rica Bruinsma, Bryan Marks, Mark Shannon, Dave Ruff, & Barry Schweitzer

Life Technologies, 501 Charmany Drive, Madison, WI, USA, 53719

Protein post-translational modifications (PTM) and protein-protein interactions (PPI) contribute to the regulation of cellular signaling networks by influencing the function, stability, and localization of proteins. As a result, monitoring changes in select PTMs and PPIs as part of the drug discovery process is important. Although numerous methods exist to measure PTMs or PPIs, most are labor intensive and time-consuming. We have applied quantitative TaqMan® protein assays, which are based on the Proximity Ligation Assay (PLATM), to measure changes in PTMs and PPIs as it offers the following advantages: limited sample requirement, homogeneous assay, robust signal amplification via TaqMan® real-time PCR, and quantitative results within a few hours. We have successfully applied PLA to monitor PTM & PPI changes to the transcription factor p53 within cells subjected to genotoxic stress. Due to the specificity, sensitivity, and small sample requirement, TaqMan® Protein Assays are an appealing alternative that can be applied at multiple stages of the drug discovery process.

 
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