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    Lysosomal Membrane Permeability (LMP) assay

    Scott Vermilyea

    Published: 2024-06-26 DOI: 10.17504/protocols.io.eq2lywxbevx9/v1
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    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.

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