Preface... 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

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

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

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

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

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

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

7. Use of the beta-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

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

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

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

Part IV Interrogation of Calpastatin, The Endogenous Calpain Inhibitor

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

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

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

Part V Manipulation of Calpain Expression

1. Experimental Manipulation of Calpain Activity In Vitro Seiichi Kitagawa

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

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

4. Methods of Calpain Inhibition to Determine the Role of Calpains in Embryo Development in Amphibians Ioanna Antoniades, Anna Charalambous, Neophytos Christodoulou, Sara Zanardelli, and Paris A. Skourides

5. Genetic Models of Calpain Deficiency and Ectopic Expression Yan Gao, Christine Hall, James MacLeod, and Peter A. Greer