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Wydawnictwo Springer Nature Customer Service Center GmbH
Data wydania 01/01/2019
Wydanie Pierwsze
Liczba stron 276
Forma publikacji książka w twardej oprawie
Poziom zaawansowania Dla profesjonalistów, specjalistów i badaczy naukowych
Język angielski
ISBN 9781493989874
Kategorie Białka
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$208.97 / €171.01 / £147.42 /
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Opis książki

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 more about this developing field, and get inspired to make new discoveries that will aid in diagnosing and treating calpain-related diseases.


Spis treści

Table of Contents...
Contributing Authors...

Part I Production and Purification of Calpains

1. Single-Step Purification of Calpain-1, Calpain-2, and Calpastatin using Anion Exchange Chromatography
Ashim Kumar Biswas and Samarth Tandon

2. Bacterial Expression and Purification of Calpains
Christian-Scott E. McCartney and Peter L. Davies

Part II Determination of Calpain Localization, Expression, and Activity

3. Casein Zymography for Analysis of Calpain-1 and Calpain-2 Activity
Ashim Kumar Biswas and Samarth Tandon

4. FRET-Based Assays to Determine Calpain Activity
Christian-Scott E. McCartney and Peter L. Davies

5. In Vivo Monitoring of Calpain Activity by Forster Resonance Energy Transfer
William Lostal, Daniel Stockholm, and Isabelle Richard

6. Determining Temporal and Spatial Expression of Calpains in Amphibians
Anna Charalambous, Ioanna Antoniades, Neophytos Christodoulou, Sara Zanardelli, and Paris A. Skourides

7. Immunohistochemical Localization of Calpains in the Amphibian Xenopus laevis
Jean-Marie Exbrayat, Elara N. Moudilou, and Claire Brun

8. Expression and Activity of Calpain A in Drosophila melanogaster
Maira Cardoso, Danielle Oliveira, and Helena Araujo

9. Use of the -Glucoronidase Reporter System (GUS) to Localize Promoter Activities of the Endogenous Plant Calpain DEFECTIVE KERNEL1 (DEK1)
Zhe Liang and Hilde-Gunn Opsahl-Sorteberg

Part III Identification of Calpain Substrates and Calpain-Activated Protein Functions

10. LabCaS for Ranking Potential Calpain Substrate Cleavage Sites from Amino Acid Sequence
Yong-Xian Fan, Xiaoyong Pan, Yang Zhang, and Hong-Bin Shen

11. CalCleaveMKL: A Tool for Calpain Cleavage Prediction
David A. duVerle and Hiroshi Mamitsuka

12. Identification of Calpain-Activated protein Functions
Maria del Carmen Lafita-Navaro and Maralice Conacci-Sorrell

Part IV Interrogation of Calpastatin, The Endogenous Calpain Inhibitor

13. Production and Purification of Recombinant Calpastatin
R. De Tullio and M. Averna

14. Isolation of Endogenous Calpastatin
R. De Tullio and M. Averna

15. Immunoblotting for Calpastatin Expression
M. Averna and R. De Tullio

Part V Manipulation of Calpain Expression

16. Experimental Manipulation of Calpain Activity In Vitro
Seiichi Kitagawa

17. In Vivo Calpain Knockdown using Delivery of siRNA
Xiaoqian Liu and Guochang Hu

18. In Vivo Expression of Mutant Calpains in the Eye using Lentivirus
Katherine J. Wert and Vinit B. Mahajan

19. Methods of Calpain Inhibition to Determine the Role of Calpains in Embryo Development in Amphibian

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