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Calpain pp 195-206 | Cite as

Immunoblotting for Calpastatin Expression

  • Monica AvernaEmail author
  • Roberta De Tullio
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1915)

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.

Key words

Immunoblot Calpastatin Calpain Protein elution Ca2+-dependent proteolysis Calpastatin phosphorylation 

Notes

Acknowledgment

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

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Experimental Medicine (DIMES)—Biochemistry SectionUniversity of GenovaGenovaItaly
  2. 2.Centre of Excellence for Biomedical Research (CEBR)University of GenovaGenovaItaly

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