Quantification of Total Human Alpha-1 Antitrypsin by Sandwich ELISA
UMass Chan Affiliations
Department of Pediatrics, Division of Pulmonary and AllergyHorae Gene Therapy Center
Document Type
Book ChapterPublication Date
2017-07-28Keywords
AATAlpha-1 antitrypsin
Enzyme-linked immunosorbent assay
Sandwich ELISA
Molecular Biology
Research Methods in Life Sciences
Metadata
Show full item recordAbstract
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.Source
Methods Mol Biol. 2017;1639:211-216. doi: 10.1007/978-1-4939-7163-3_20. Link to article on publisher's siteDOI
10.1007/978-1-4939-7163-3_20Permanent Link to this Item
http://hdl.handle.net/20.500.14038/43592PubMed ID
28752460Related Resources
Link to Article in PubMedae974a485f413a2113503eed53cd6c53
10.1007/978-1-4939-7163-3_20