DQ-BSA assay
Narayana Yadavalli, Shawn M. Ferguson
Published: 2023-07-08 DOI: 10.17504/protocols.io.kxygx9r9dg8j/v1
Abstract
This protocol describes the DQ BSA assay for measuring the lysosomal proteolytic activity.
Attachments
Steps
DQ-BSA assay
1.
Plate 100,000 mature macrophages or microglia or BMDM were seeded on Mattek glass bottom dishes.
2.
Next day treat cells with 50nanomolar (nM) MLi2 or 250nanomolar (nM) LRRK2-IN-1 for 2h 0m 0s followed by 1h 0m 0s treatment with 10µg/mL DQ-BSA (Thermo Fisher Scientific, #D12051) and 50µg/mL Alexa-488 BSA (Thermo Fisher Scientific #A13100) in the media containing inhibitors.
3.
After 3 hours wash the cells 3X with culture media.
4.
Then add 1mL culture media to the washed cells and return to incubator for 0h 10m 0s.
5.
Imaging was done on LSM 880.
