Full Signaling Pathway Analysis In A Single Run

Gaining an accurate quantitative understanding of complex signaling networks as a whole is crucial and remains a daunting challenge. Because signaling networks are complex, a high-throughput method is often needed to measure all protein targets involved, including phosphorylated and non-phosphorylated protein forms of each protein in the signaling cascade. Traditional Western blots are commonly chosen for signaling studies over other immunoassay approaches since the provided molecular weight information paired with immunodetection increases assay specificity, which can be a challenge when looking at phosphorylated protein isoforms or other post translational modifications. Nonetheless, traditional Western blots are typically limited in their throughput and ability to profile more than 1 or 2 proteins in the signaling pathway at a time.

Here, we show how the high throughput and low sample volume requirements of Simple Western work together to comprehensively characterize complex signaling pathways in one simple workflow.