Alpha-1-antitrypsin deficiency : methods and protocols

Description

This volume provides protocols that expand on the latest alpha-1-antitrypsin (AAT) research. The chapters in this book are divided in to three sections: Part I is dedicated to patient-oriented research; part II discusses animal models; and Part III focuses on in vitro studies. 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 authoritative, Alpha-1 Antitrypsin Deficiency: Methods and Protocols is a valuable resource for researchers, students, and clinican-scientists interested in AAT deficiency, as well as anyone working in the fields of pulmonology and hepatology. Front Matter ....Pages i-xiii Pathophysiology of Alpha-1 Antitrypsin Deficiency Liver Disease (Jeffrey H. Teckman, Keith S. Blomenkamp)....Pages 1-8 Pathophysiology of Alpha-1 Antitrypsin Lung Disease (Michael Kalfopoulos, Kaitlyn Wetmore, Mai K. ElMallah)....Pages 9-19 Measuring and Interpreting Serum AAT Concentration (Leslie J. Donato, Melissa R. Snyder, Dina N. Greene)....Pages 21-32 AAT Phenotype Identification by Isoelectric Focusing (Dina N. Greene, M. C. Elliott-Jelf, David G. Grenache)....Pages 33-44 Laboratory Diagnosis by Genotyping (Irene Belmonte, Luciana Montoto, Francisco Rodríguez-Frías)....Pages 45-60 Genotyping Protocol for the Alpha-1 Antitrypsin (PiZ) Mouse Model (Alisha M. Gruntman)....Pages 61-65 Elastase-Induced Lung Emphysema Models in Mice (Béla Suki, Erzsébet Bartolák-Suki, Patricia R. M. Rocco)....Pages 67-75 Assessing Structure–Function Relations in Mice Using the Forced Oscillation Technique and Quantitative Histology (Harikrishnan Parameswaran, Béla Suki)....Pages 77-91 Practical Methods for Assessing Emphysema Severity Based on Estimation of Linear Mean Intercept (Lm) in the Context of Animal Models of Alpha-1 Antitrypsin Deficiency (Airiel M. Davis, Kristen E. Thane, Andrew M. Hoffman)....Pages 93-106 Design, Cloning, and In Vitro Screening of Artificial miRNAs to Silence Alpha-1 Antitrypsin (Florie Borel, Christian Mueller)....Pages 107-114 Methods to Identify and Characterize siRNAs Targeting Serpin A1 In Vitro and In Vivo Using RNA Interference (Stuart Milstein, Kun Qian, Linying Zhang, Alfica Sehgal)....Pages 115-126 Knockdown of Z Mutant Alpha-1 Antitrypsin In Vivo Using Modified DNA Antisense Oligonucleotides (Mariam Aghajan, Shuling Guo, Brett P. Monia)....Pages 127-138 Immunohistochemistry Staining for Human Alpha-1 Antitrypsin (Dongtao A. Fu, Martha Campbell-Thompson)....Pages 139-143 Periodic Acid-Schiff Staining with Diastase (Dongtao A. Fu, Martha Campbell-Thompson)....Pages 145-149 Protocol for Directed Differentiation of Human Induced Pluripotent Stem Cells (iPSCs) to a Hepatic Lineage (Joseph E. Kaserman, Andrew A. Wilson)....Pages 151-160 Isolation of Kupffer Cells and Hepatocytes from a Single Mouse Liver (Marcela Aparicio-Vergara, Michaela Tencerova, Cecilia Morgantini, Emelie Barreby, Myriam Aouadi)....Pages 161-171 Alpha-1 Antitrypsin Transcytosis and Secretion (Angelia D. Lockett)....Pages 173-184 Measuring the Effect of Histone Deacetylase Inhibitors (HDACi) on the Secretion and Activity of Alpha-1 Antitrypsin (Chao Wang, Marion Bouchecareilh, William E. Balch)....Pages 185-193 Expression and Purification of Active Recombinant Human Alpha-1 Antitrypsin (AAT) from Escherichia coli (Beena Krishnan, Lizbeth Hedstrom, Daniel N. Hebert, Lila M. Gierasch, Anne Gershenson)....Pages 195-209 Quantification of Total Human Alpha-1 Antitrypsin by Sandwich ELISA (Qiushi Tang, Alisha M. Gruntman, Terence R. Flotte)....Pages 211-216 Quantification of Murine AAT by Direct ELISA (Andrew Cox, Christian Mueller)....Pages 217-222 Quantification of Z-AAT by a Z-Specific “Sandwich” ELISA (Florie Borel, Qiushi Tang, Christian Mueller)....Pages 223-226 Semiquantitation of Monomer and Polymer Alpha-1 Antitrypsin by Centrifugal Separation and Assay by Western Blot of Soluble and Insoluble Components (Keith S. Blomenkamp, Jeffrey H. Teckman)....Pages 227-234 Electrophoresis- and FRET-Based Measures of Serpin Polymerization (Sarah V. Faull, Anwen E. Brown, Imran Haq, James A. Irving)....Pages 235-248 Therapeutics: Alpha-1 Antitrypsin Augmentation Therapy (Michael Campos, Jorge Lascano)....Pages 249-262 Therapeutic Options in Alpha-1 Antitrypsin Deficiency: Liver Transplantation (Nedim Hadzic)....Pages 263-265 Therapeutics: Gene Therapy for Alpha-1 Antitrypsin Deficiency (Alisha M. Gruntman, Terence R. Flotte)....Pages 267-275 Back Matter ....Pages 277-278