Jun 25, 2024
Lysosomal Membrane Permeability (LMP) assay
- Scott Vermilyea1
- 1University of Minnesota
Protocol Citation: Scott Vermilyea 2024. Lysosomal Membrane Permeability (LMP) assay. protocols.io https://dx.doi.org/10.17504/protocols.io.eq2lywxbevx9/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: June 07, 2024
Last Modified: June 25, 2024
Protocol Integer ID: 102395
Keywords: Lysosomal Membrane Permeability (LMP) assay, Antibody, Cell culture, Imaging
Funders Acknowledgement:
ASAP
Grant ID: 000592
Abstract
This protocol details the requirements of the Lysosomal Membrane Permeability (LMP) assay.
Materials
• Antibody:
Anti-Galectin 3 antibody (ab2785)Anti-Galectin 3 antibody [A3A12] AbcamCatalog #ab2785
• Coverslips:
Poly-D-Lysin coated coverslips. Corning BioCoat (Ref: 354086).
Corning® BioCoat™ Poly-D-Lysine #1 German Glass CoverslipMerckCatalog #354086
Cell Culture/Treatments:
Cell Culture/Treatments:
0.5x10^4/ml of cells plated on coverslips.
PFF-488 treatment.
PBS/trypsin (0.01%) wash.
Fix cells using 4% PFA.
Immunostaining using Galectin 3 primary antibody.
Imaging:
Imaging:
Leica Stellaries 8 confocal laser scanning microscope.
Image analysis:
Image J. quantification of the % area of the cell covered by Galectin 3 immunoreactivity.