| First Author | Frost LS | Year | 2017 |
| Journal | Methods Enzymol | Volume | 587 |
| Pages | 43-54 | PubMed ID | 28253971 |
| Mgi Jnum | J:260859 | Mgi Id | MGI:6148146 |
| Doi | 10.1016/bs.mie.2016.09.052 | Citation | Frost LS, et al. (2017) The Use of DQ-BSA to Monitor the Turnover of Autophagy-Associated Cargo. Methods Enzymol 587:43-54 |
| abstractText | There is increasing evidence documenting the critical role played by autophagic and autophagy-associated processes in maintaining cell homeostasis and overall systemic health. Autophagy is considered a degradative as well as a recycling pathway that relies on encapsulated intracellular components trafficking to and fusing with degradative compartments, including lysosomes. In this chapter, we describe the use of DQ-BSA to study autophagosome-lysosome fusion as well as a means by which to analyze hybrid autophagic pathways. Such noncanonical pathways include LC3-associated phagocytosis, better known as LAP. Both autophagosomes and LAPosomes (LC3-associated phagosomes) deliver cargo for degradation. The use of fluorescent DQ-BSA in conjugation with autophagic makers and biomarkers of hybrid autophagy offers a reliable technique to monitor the formation of autolysosomes and LAPo-lysosomes in both fixed- and live-cell studies. This technique relies on cleavage of the self-quenched DQ Green- or DQ Red BSA protease substrates in an acidic compartment to generate a highly fluorescent product. |
