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Isolation of Endogenous Calpastatin

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Book cover Calpain

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

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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.

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References

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Acknowledgments

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

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Authors and Affiliations

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

    Roberta De Tullio & Monica Averna

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

    Roberta De Tullio

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

Correspondence to Roberta De Tullio .

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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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De Tullio, R., Averna, M. (2019). Isolation of Endogenous Calpastatin. 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_14

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  • DOI: https://doi.org/10.1007/978-1-4939-8988-1_14

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  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-4939-8987-4

  • Online ISBN: 978-1-4939-8988-1

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