Autophagy or autophagocytosis ("self-eating") refers to a process of degradation of cytoplasmic components within lysosomes, a unique process mediated by autophagosomes. Autophagy consists of several sequential steps: sequestration, transport to lysosomes, degradation and eventual re-utilization of degradation products. Autophagosomes engulf a portion of cytoplasm, thus autophagy is generally thought to be a nonselective degradation system regulated by many different cellular signaling pathways. Autophagy functions as a stress response upregulated by nutrient and energy starvation, oxidative stress, or other harmful conditions (such as damage to organelles, protein aggregation and infection by pathogens). Dysfunction of autophagy is associated with many human cancers and neurodegenerative diseases. Assay Genie's Autophagy/Cytotoxicity Dual Staining Kit (Cell-Based) enables detection and monitoring of autophagy and autophagic cell death in cultured cells. The kit contains two fluorescent probes: a membrane permeable selective autophagy stain and a cell death marker. An autophagy-inducing positive control reagent, which increases autophagy staining and serves as an experimental control. This easy-to-use non-radioactive kit allows researchers to study the regulation of autophagy and cytotoxicity at the cellular level by Fluorescence Microscopy and Flow Cytometry in cultured cells.

Figure: Autophagy/Cytotoxicity Staining in Jurkat and HeLa cells. 1x10 6 Jurkat or 1x10 5 HeLa cells were treated with vehicle or Positive Control Reagent for 24 h followed by Autophagy Staining according to kit’s protocol. Histograms from flow analysis of untreated cells (Basal Autophagy; A) and Experimental Control Cells (treated with Positive Control; B). Fluorescence microscope images of Basal Autophagy C) and Experimental Control Cells D). Increased blue fluorescence induced by treatment with Positive Control Reagent compared to the Basal Autophagy levels confirms intracellular accumulation of fluorescent stain in the autophagosomes.