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Calpain pp 187-194 | Cite as

Isolation of Endogenous Calpastatin

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

Abstract

We here describe the purification of calpastatin from human erythrocytes. When calpastatin is purified from tissues, it is necessary to measure its inhibitory activity against calpain in the presence of Ca2+ to specifically identify the protein. Thus, the purification steps necessary to obtain the inhibitor protein were originally designed to obtain calpain from the same tissue. For this reason, in addition to calpastatin purification, we also include a method for purifying human erythrocyte calpain and globin. We routinely use these two components for assaying calpastatin inhibition.

Key words

Calpastatin isolation Calpain inhibition Chromatography Calpain assay Human erythrocytes 

Notes

Acknowledgments

This work was supported by the grant FRA2015 and FRA2016 from the University of Genova 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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