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This volume provides a collection of protocols that will allow researchers to further explore the multiple, fascinating activities of Alpha-1 Antitrypsin (AAT) isoforms in innate immunity, and gene therapy. 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.
Authoritative and cutting-edge, Alpha-1 Antitrypsin: Methods and Protocols aims to ensure successful results in the further study of this vital field.
List of contents
Introduction: Alpha-1 Antitrypsin Deficiency.- Approaches to Therapeutic Gene Editing in Alpha-1 Antitrypsin Deficiency.-Manipulation of Proteostasis Networks in Transgenic ZAAT Zebrafish via CRISPR-Cas9 Gene Editing.- PCR Analysis to Identify AAT Gene Promoters and Splice Variants.- Native and Ion Mobility Mass Spectrometry Characterization of Alpha-1 Antitrypsin Variants and Oligomers.- Sanger and Next Generation Sequencing of AAT.- Measuring the Concentration of Active Alpha-1 Antitrypsin in Serum.- Characterization of Novel Alpha-1 Antitrypsin Coding Variants in a Mammalian Cellular Model.- Alpha-1 Antitrypsin (A1AT) Proteotyping by LC-MS/MS.- Serum Western Blot for the Detection of a c-Myc Protein Tag in Non-Human Primates and Mice.- An Enzyme-Linked Immunosorbent Assay (ELISA) for Quantification of Circulating Pi*Z Alpha-1 Antitrypsin Polymers.- Measuring of Alpha1 Antitrypsin Concentration by Nephelometry or Turbidimetry.- Autoantibody Recognition of Natural and Homocysteinylated Alpha-1 Antitrypsin: Indirect Enzyme-Linked ImmunoSorbent Assay (ELISA) Quantification in Sera.- Monitoring the Secretion and Activity of Alpha-1 Antitrypsin in Various Mammalian Cell Types.- Lipoproteins in Negative Feedback with Alpha-1 Antitrypsin.- Silencing Very Low Density Lipoprotein Receptor Reveals Alpha-1 Antitrypsin Role in HIV infectivity.- Purified versus Plasma Alpha-1 Antitrypsin Effects on Cellular Activities.
