The measurement of autophagic flux is critical in understanding the regulation of autophagy.
The Pho8Δ60 assay employs a very sensitive enzymatic assay that provides a high signal‐
to‐noise ratio and allows for precise quantification of autophagic flow in yeast. Pho8,
alkaline phosphatase, is a resident vacuolar enzyme that is delivered to the vacuole
membrane through a portion of the secretory pathway. The assay utilizes a genetically
engineered version of Pho8 that lacks the N‐terminal transmembrane domain that allows for …