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.

Includes cutting-edge methods and protocols.

Pathophysiology of Alpha-1 Antitrypsin Deficiency Liver Disease.- Pathophysiology of Alpha-1 Antitrypsin Lung Disease.- Measuring and Interpreting Serum AAT Concentration.- AAT Phenotype Identification by Isoelectric Focusing.- Laboratory Diagnosis by Genotyping.- Genotyping Protocol for the Alpha-1 Antitrypsin (PiZ) Mouse Model.- Elastase-Induced Lung Emphysema Models in Mice.- Assessing Structure-Function Relations in Mice Using the Forced Oscillation Technique and Quantitative Histology.- 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.- Design, Cloning, and In Vitro Screening of Artificial miRNAs to Silence Alpha-1-Antitrypsin.- Methods to Identify and Characterize siRNAs Targeting Serpin A1 In Vitro and In Vivo using RNA Interference.- Knockdown of Z Mutant Alpha-1 Antitrypsin in Vivo Using Modified DNA Antisense Oligonucleotides.- Immunohistochemistry Staining for Human Alpha-1 Antitrypsin.- Periodic Acid-Schiff Staining with Diastase.- Protocol for Directed Differentiation of Human Induced Pluripotent Stem Cells (iPSCs) to a Hepatic Lineage.- Isolation of Kupffer Cells and Hepatocytes from a Single Mouse Liver.- Alpha-1 Antitrypsin Transcytosis and Secretion.- Measuring the Effect of Histone Deacetylase Inhibitors (HDACi) on the Secretion and Activity of Alpha-1 Antitrypsin.- Expression and Purification of Active Recombinant Human ¿1-Antitrypsin (AAT) from Escherichia coli.- Quantification of Total Human Alpha-1 Antitrypsin by Sandwich ELISA.- Quantification of Murine AAT by Direct ELISA.- Quantification of Z-AAT by a Z-Specific "Sandwich" ELISA.- Semi-Quantitation of Monomer and Polymer Alpha-1 Antitrypsin by Centrifugal Separation and Assay by Western Blot of Soluble and Insoluble Components.- Electrophoresis- and FRET-Based Measures of Serpin Olymerization.- Therapeutics: Alpha-1 Antitrypsin Augmentation Therapy.- Therapeutic Option in Alpha-1 Antitrypsin Liver Transplantation.- Therapeutics: Gene Therapy for Alpha-1 Antitrypsin Deficiency.