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Immunoblotting for Calpastatin Expression

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Calpain

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1915))

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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.

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Acknowledgment

This work was supported by grants FRA2015 and FRA2016 from the University of Genoa to MA and RDT.

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Authors and Affiliations

  1. Department of Experimental Medicine (DIMES)—Biochemistry Section, University of Genova, Genova, Italy

    Monica Averna & Roberta De Tullio

  2. Centre of Excellence for Biomedical Research (CEBR), University of Genova, Genova, Italy

    Roberta De Tullio

Authors
  1. Monica Averna
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  2. Roberta De Tullio
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Corresponding author

Correspondence to Monica Averna .

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Editors and Affiliations

  1. Department of Inflammation and Immunity, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA

    Jeannette S. Messer

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Averna, M., De Tullio, R. (2019). Immunoblotting for Calpastatin Expression. In: Messer, J. (eds) Calpain. Methods in Molecular Biology, vol 1915. Springer, New York, NY. https://doi.org/10.1007/978-1-4939-8988-1_15

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

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  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-4939-8987-4

  • Online ISBN: 978-1-4939-8988-1

  • eBook Packages: Springer Protocols

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