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.
CITATION STYLE
Tang, Q., Gruntman, A. M., & Flotte, T. R. (2017). Quantification of total human alpha-1 antitrypsin by sandwich ELISA. In Methods in Molecular Biology (Vol. 1639, pp. 211–216). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7163-3_20
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