Expression and Activity of Calpain A in Drosophila melanogaster
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.
Key wordsCalpain A Drosophila Calpain activity Fluorogenic substrate
This work was supported by CNPq/Brazil grant to HA and fellowship to MAC.
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