Lysosomal activity DQ-Red BSA assay
Razaul Karim
Published: 2024-06-26 DOI: 10.17504/protocols.io.j8nlk8dp6l5r/v1
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Abstract
This protocol details the Lysosomal activity DQ-Red BSA assay.
Steps
Cell Culture/Treatments:
1.
0.5x10^4/ml of cells plated on coverslips.
2.
PFF-488 treatment.
3.
PBS/trypsin (0.01%) wash.
4.
DQ-BSA Red treatment:
10``1h 30m 0s at 37°C.
5.
Fix cells using 4% PFA.
Imaging:
6.
Red fluorescence signal is detected only when fluorogenic substrate is hydrolyzed by lysosomal proteases.
7.
Leica Stellaries 8 confocal laser scanning microscope.
8.
Image analysis:
Image J. quantification of the % area of the cell covered by DQ-Red signal divided by the number of cells (DAPI stained).
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