Title

Quantification of Total Human Alpha-1 Antitrypsin by Sandwich ELISA

UMMS Affiliation

Horae Gene Therapy Center; Department of Pediatrics, Division of Pulmonary and Allergy

Date

7-28-2017

Document Type

Book Chapter

Disciplines

Molecular Biology | Research Methods in Life Sciences

Abstract

In this chapter we describe an enzyme-linked immunosorbent assay (ELISA) to quantitatively measure human alpha-1 antitrypsin (AAT) protein levels in serum, other body fluids or liquid media. This assay can be used to measure the expression of the human AAT (hAAT) gene in a variety of gene transfer or gene downregulation experiments.A hAAT-specific capture antibody and a HRP-conjugated anti-AAT detection antibody are used in this assay. The conjugated anti-AAT used in this protocol, instead of the typical sandwich which employs an unconjugated antibody followed by a specifically conjugated IgG, makes the assay simpler and decreases variability. This provides a useful tool to evaluate the AAT levels in clinical and research samples and can allow fairly rapid testing of a large number of samples.

Rights and Permissions

Citation: Methods Mol Biol. 2017;1639:211-216. doi: 10.1007/978-1-4939-7163-3_20. Link to article on publisher's site

Related Resources

Link to Article in PubMed

Keywords

AAT, Alpha-1 antitrypsin, Enzyme-linked immunosorbent assay, Sandwich ELISA

PubMed ID

28752460