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.