Mammalian beta-Galactosidase Assay Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Add 100µL of β-Galactosidase Assay Reagent. Cover plate and incubate for 30 minutes at 37°C.	Measure absorbance at 405nm.

Protocol tips
Add 100µL of β-Galactosidase Assay Reagent. Cover plate and incubate for 30 minutes at 37°C.
Downstream tips
Measure absorbance at 405nm.

Publication protocol

For lysosomal enzyme activities, live CMs were incubated with an enzymatic substrate for 1 h at 37°C and the product formation was assessed by fluorescence microscopy. For CatL activity, CMs were incubated with MR-CatL substrate (Immunochemistry Technologies, Bloomington, MN). For lysosomal acid phosphatase (LAP) activity, CMs were incubated with LysoLive Phosgreen substrate (Marker Gene Technologies, Eugene, OR). Images were acquired as described in the supplementary Methods section. The lysosomal fraction described earlier was used to measure β-galactosidase activity using the mammalian β-galactosidase assay kit (Thermo Scientific) and β-hexosaminidase activity as previously reported (25).

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