Magic Red Cathepsin B activity assay
Razaul Karim
Published: 2024-06-26 DOI: 10.17504/protocols.io.ewov192b2lr2/v1
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Abstract
This protocol details the magic red Cathepsin B activity 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.
1x Magic Red solution for last 0h 15m 0s at 37°C in cell culture incubator.
Imaging
5.
Live-Cell imaging.
6.
Leica Stellaries 8 confocal laser scanning microscope, assembled with live cell chamber with CO2 gas, temperature and humidity controlled.
7.
Image analysis:
Image J. quantification of the % area of the cell covered by Magic Red signal, as well as total signal intensity per cell.
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