Expression and Activity of Calpain A in Drosophila melanogaster.
CALPAIN: METHODS AND PROTOCOLS(2019)
摘要
Detecting calpain activity in Drosophila tissues is a fundamental tool to study calpain function. We use differential centrifugation to prepare membrane- versus cytosol-enriched fractions for measuring calpain activity with the fluorogenic substrate N-LY-AMC. With this method one can measure calpain A activity in wild-type flies and in several mutant fly backgrounds, revealing a strong correlation between in situ membrane distribution and in vitro determined activity measurements. Here we describe the steps for tissue preparation and calpain activity measurement in the Drosophila embryo.
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关键词
Calpain A, Drosophila, Calpain activity, Fluorogenic substrate
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