Alpha-1 Antitrypsin, 1st ed. 2024 Methods and Protocols Methods in Molecular Biology Series, Vol. 2750

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.

1)      Introduction: Alpha-1 Antitrypsin Deficiency

Alisha M. Gruntman, Wen Xue, Terence R. Flotte

Part I: Alpha-1 Antitrypsin polymorphisms and gene exon usage

2)      Approaches to Therapeutic Gene Editing in Alpha-1 Antitrypsin Deficiency

Alisha M. Gruntman, Wen Xue, and Terence R. Flotte

3)      Manipulation of Proteostasis Networks in Transgenic ZAAT Zebrafish via CRISPR-Cas9 Gene Editing

Connie Fung, Lee B. Miles, Robert J. Bryson-Richardson, and Phillip I. Bird

4)      PCR Analysis to Identify AAT Gene Promoters and Splice Variants

Noor Kalsheker

5)      Native and Ion Mobility Mass Spectrometry Characterization of Alpha-1 Antitrypsin Variants and Oligomers

Sarah Vickers, James Irving, David A. Lomas, and Konstantinos Thalassinos

6)      Sanger and Next Generation Sequencing of AAT

Valentina Barzon, Ilaria Ferrarotti, and Stefania Ottaviani

Part II: AAT protein levels and specific activity

7)      Measuring the Concentration of Active Alpha-1 Antitrypsin in Serum

Cynthia L. Bristow

Andrea Denardo, Emna Ben Khlifa, Mattia Bignotti, and Annamarie Fra

9)      Alpha-1 Antitrypsin (A1AT) Proteotyping by LC-MS/MS

Jennifer Kemp, Paula M. Ladwig, and Melissa Snyder

10)   Serum Western Blot for the Detection of a c-Myc Protein Tag in Non-Human Primates and Mice

Meghan Blackwood, Qiushi Tang, and Alisha M. Gruntman

11)   An Enzyme-Linked Immunosorbent Assay (ELISA) for Quantification of Circulating Pi*Z Alpha-1 Antitrypsin Polymers

Sabina Janciauskiene, Tobias Welte, and Matthias Lehmann

12)   Measuring of Alpha1 Antitrypsin Concentration by Nephelometry or Turbidimetry

13)   Autoantibody Recognition of Natural and Homocysteinylated Alpha-1 Antitrypsin: Indirect Enzyme-Linked ImmunoSorbent Assay (ELISA) Quantification in Sera

Tania Colasanti and Fabrizio Conti

14)   Monitoring the Secretion and Activity of Alpha-1 Antitrypsin in Various Mammalian Cell Types

Kevin P. Guay, Haiping Ke, Lila M. Gierasch, Anne Gershenson, and Daniel N. Hebert

Part III: AAT interactions with lipoproteins and lipoprotein receptors

15)   Lipoproteins in Negative Feedback with Alpha-1 Antitrypsin

Cynthia L. Bristow

16)   Silencing Very Low Density Lipoprotein Receptor Reveals Alpha-1 Antitrypsin Role in HIV infectivity

Cynthia L. Bristow

17)   Purified versus Plasma Alpha-1 Antitrypsin Effects on Cellular Activities

Cynthia L. Bristow

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts