Quantification of Total Human Alpha-1 Antitrypsin by Sandwich ELISA
Horae Gene Therapy Center; Department of Pediatrics, Division of Pulmonary and Allergy
Molecular Biology | Research Methods in Life Sciences
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.
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Citation: Methods Mol Biol. 2017;1639:211-216. doi: 10.1007/978-1-4939-7163-3_20. Link to article on publisher's site
AAT, Alpha-1 antitrypsin, Enzyme-linked immunosorbent assay, Sandwich ELISA
Tang, Qiushi; Gruntman, Alisha; and Flotte, Terence R., "Quantification of Total Human Alpha-1 Antitrypsin by Sandwich ELISA" (2017). Pediatric Publications and Presentations. 162.