Immunoblotting for Calpastatin Expression.

Monica Averna, Roberta De Tullio

Journal: Methods in molecular biology (Clifton, N.J.) 2019;1915():195-206

PMID: 30617805

Abstract

Immunoblotting is a procedure routinely used to analyze calpastatin expression. However, immunoblotting alone may not be adequate for this task, since calpastatin isoforms can vary by tissue, can be modified by partial digestion, and can undergo posttranslational modifications. Here we describe a method for more precise evaluation of calpastatin expression by combining immunoblot analysis with an assay for the inhibitory activity of a single calpastatin species isolated by SDS-PAGE and protein elution from the gel.

Address: Department of Experimental Medicine (DIMES)-Biochemistry Section, University of Genova, Genova, Italy. [email protected].; Department of Experimental Medicine (DIMES)-Biochemistry Section, University of Genova, Genova, Italy.; Centre of Excellence for Biomedical Research (CEBR), University of Genova, Genova, Italy.

DOI: 10.1007/978-1-4939-8988-1_15

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https://doi.org/10.1007/978-1-4939-8988-1_15

MeSH Terms: Calcium-Binding Proteins, Calpain, Electrophoresis, Polyacrylamide Gel, Gene Expression, Immunoblotting, Muscles