I am trying to standardise lysosomal enzymes assay. Since the enzyme is active inside the lysosome and is partially embedded in the lysosomal membrane, lysis of cells with lysosomal membrane is crucial. I'm trying to standardise cycles and timing to get proper enzyme assay after cell lysis. Have anyone tried sonication for lysosomal enzymes?
Hi Dipti Deshpande A long time ago I was measuring lysosomal enzyme activities in cell lysates. You really don't need to use hard sonication like for bacteria to disrupt the lysosomes.
Pipetting the lysates up/down with a small bore syringe needle in the presence of mild detergents (0.1% tween, TX100) or sonicating using a jewelry waterbath sonicator for ~5 min in the presence of the detergents will dissolve any eukaryotic lipid structures except for mitochondria and some nuclei.
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