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Application Note

Detection and Quantification of Autophagy Using the AlphaLISA p62 Assay

Introduction

Lysosomal dysregulation is the hallmark of many diseases including neurodegenerative diseases, lysosomal storages diseases, and aging. The lysosome is responsible for eliminating cellular waste in a multistep pathway called autophagy. This process is well regulated and can be affected by a number of stimulants including nutrient starvation, physiological stress, and chemical induction. Sequestosome-1, also known as p62, is incorporated into autophagosomes then subsequently degraded in the final steps of autophagy. Assessing this protein can help decipher a block or increased flux in autophagy.

Quantification of p62 levels within cellular lysates is often performed with labor intensive wash-based ELISA assays. In this study, chloroquine is used as an inhibitor of autophagy to show how the homogeneous no-wash AlphaLISA® p62 assay can detect changing levels of this lysosomal protein.