Reference - Detail
| RRC ID | 47684 |
|---|---|
| Author | Frost LS, Dhingra A, Reyes-Reveles J, Boesze-Battaglia K. |
| Title | The Use of DQ-BSA to Monitor the Turnover of Autophagy-Associated Cargo. |
| Journal | Methods Enzymol |
| Abstract |
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. |
| Volume | 587 |
| Pages | 43-54 |
| Published | 2016-11-17 |
| DOI | 10.1016/bs.mie.2016.09.052 |
| PII | S0076-6879(16)30325-1 |
| PMID | 28253971 |
| PMC | PMC5338641 |
| MeSH | Autophagosomes / metabolism Autophagy* Boron Compounds / chemistry Cell Line Epithelial Cells / metabolism Fluorescent Dyes / chemistry Fluorescent Dyes / metabolism Humans Lysosomes / metabolism Microscopy, Confocal / methods Microtubule-Associated Proteins / analysis Microtubule-Associated Proteins / metabolism* Molecular Biology / methods* Serum Albumin, Bovine / chemistry Serum Albumin, Bovine / metabolism* |
| IF | 1.394 |
| Times Cited | 17 |
| Resource | |
| Mice | RBRC00806 |