Lysosomal Membrane Permeability (LMP) assay
Scott Vermilyea
Published: 2024-06-26 DOI: 10.17504/protocols.io.eq2lywxbevx9/v1
Abstract
This protocol details the requirements of the Lysosomal Membrane Permeability (LMP) 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.
Fix cells using 4% PFA.
5.
Immunostaining using Galectin 3 primary antibody.
Imaging:
6.
Leica Stellaries 8 confocal laser scanning microscope.
7.
Image analysis:
Image J. quantification of the % area of the cell covered by Galectin 3 immunoreactivity.
