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Calpain pp 67-79 | Cite as

Determining Temporal and Spatial Expression of Calpains in Amphibians

  • Anna Charalambous
  • Ioanna Antoniades
  • Neophytos Christodoulou
  • Sara Zanardelli
  • Paris A. SkouridesEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1915)

Abstract

Calpains are a family of calcium-dependent intracellular cysteine proteases that regulate important physiological processes by substrate cleavage. Despite the fact that Calpains have been identified in the Xenopus genome, their expression patterns and role have not been characterized. Therefore, herein, we describe two methods to determine temporal and spatial expression of Calpain 2 during Xenopus development, namely, RT-PCR and whole-mount in situ hybridization (WISH). In addition, indirect immunofluorescence (IF) is described to determine translocation to the plasma membrane, which correlates with activity levels of Calpain 2.

Key words

Xenopus Calpain RT-PCR Whole-mount in situ hybridization Immunofluorescence 

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Copyright information

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

Authors and Affiliations

  • Anna Charalambous
    • 1
  • Ioanna Antoniades
    • 1
  • Neophytos Christodoulou
    • 1
  • Sara Zanardelli
    • 1
  • Paris A. Skourides
    • 1
    Email author
  1. 1.Department of Biological SciencesUniversity of CyprusNicosiaCyprus

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