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Calpain

Methods and Protocols

  • Book
  • © 2019

Overview

Editors:
  1. Jeannette S. Messer

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

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  • Includes cutting-edge methods and protocols
  • Provides step-by-step detail essential for reproducible results
  • Contains key notes and implementation advice from the experts

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

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Table of contents (20 protocols)

  1. Front Matter

    Pages i-xii
    Download chapter PDF

  2. Production and Purification of Calpains

    1. Front Matter

      Pages 1-1
      Download chapter PDF

    2. Single-Step Purification of Calpain-1, Calpain-2, and Calpastatin Using Anion-Exchange Chromatography

      • Ashim Kumar Biswas, Samarth Tandon
      Pages 3-11

    3. Bacterial Expression and Purification of Calpains

      • Christian-Scott E. McCartney, Peter L. Davies
      Pages 13-27

  3. Determination of Calpain Localization, Expression, and Activity

    1. Front Matter

      Pages 29-29
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    2. Casein Zymography for Analysis of Calpain-1 and Calpain-2 Activity

      • Ashim Kumar Biswas, Samarth Tandon
      Pages 31-38

    3. FRET-Based Assays to Determine Calpain Activity

      • Christian-Scott E. McCartney, Peter L. Davies
      Pages 39-55

    4. In Vivo Monitoring of Calpain Activity by Forster Resonance Energy Transfer

      • William Lostal, Daniel Stockholm, Isabelle Richard
      Pages 57-66

    5. Determining Temporal and Spatial Expression of Calpains in Amphibians

      • Anna Charalambous, Ioanna Antoniades, Neophytos Christodoulou, Sara Zanardelli, Paris A. Skourides
      Pages 67-79

    6. Immunohistochemical Localization of Calpains in the Amphibian Xenopus laevis

      • Jean-Marie Exbrayat, Elara N. Moudilou, Claire Brun
      Pages 81-92

    7. Expression and Activity of Calpain A in Drosophila melanogaster

      • Maira Cardoso, Danielle Oliveira, Helena Araujo
      Pages 93-101

    8. Use of the β-Glucuronidase (GUS) Reporter System to Localize Promoter Activities of the Endogenous Plant Calpain DEFECTIVE KERNEL1 (DEK1)

      • Zhe Liang, Hilde-Gunn Opsahl-Sorteberg
      Pages 103-108

  4. Identification of Calpain Substrates and Calpain-Activated Protein Functions

    1. Front Matter

      Pages 109-109
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    2. LabCaS for Ranking Potential Calpain Substrate Cleavage Sites from Amino Acid Sequence

      • Yong-Xian Fan, Xiaoyong Pan, Yang Zhang, Hong-Bin Shen
      Pages 111-120

    3. CalCleaveMKL: a Tool for Calpain Cleavage Prediction

      • David A. duVerle, Hiroshi Mamitsuka
      Pages 121-147

    4. Identification of Calpain-Activated Protein Functions

      • Maria del Carmen Lafita-Navarro, Maralice Conacci-Sorrell
      Pages 149-160

  5. Interrogation of Calpastatin, The Endogenous Calpain Inhibitor

    1. Front Matter

      Pages 161-161
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    2. Production and Purification of Recombinant Calpastatin

      • Roberta De Tullio, Monica Averna
      Pages 163-185

    3. Isolation of Endogenous Calpastatin

      • Roberta De Tullio, Monica Averna
      Pages 187-194

    4. Immunoblotting for Calpastatin Expression

      • Monica Averna, Roberta De Tullio
      Pages 195-206
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Keywords

About this book

This volume brings together a plethora of protocols and experimental methods used by scientists to study calpains, their inhibitors, and their substrates. It also explores bioinformatic approaches to calpain substrate identification. The chapters in this book are divided into five parts and cover topics such as production and purification of calpains; determination of calpain localization, expression, and activity; identification of calpain-activated protein function; interrogation of calpastatin; and manipulation of calpain expression. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and practical, Calpain: Methods and Protocols is a valuable resource for researchers and scientists who want to learn moreabout this developing field, and get inspired to make new discoveries that will aid in diagnosing and treating calpain-related diseases.


Editors and Affiliations

  • Department of Inflammation and Immunity, Lerner Research Institute, Cleveland Clinic, Cleveland, USA

    Jeannette S. Messer

Bibliographic Information

  • Book Title: Calpain

  • Book Subtitle: Methods and Protocols

  • Editors: Jeannette S. Messer

  • Series Title: Methods in Molecular Biology

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

  • Publisher: Springer New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media, LLC, part of Springer Nature 2019

  • Hardcover ISBN: 978-1-4939-8987-4Published: 08 January 2019

  • eBook ISBN: 978-1-4939-8988-1Published: 08 January 2019

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XII, 276

  • Number of Illustrations: 29 b/w illustrations, 37 illustrations in colour

  • Topics: Protein Science

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