Title

Quantification of Murine AAT by Direct 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

This methods chapter elaborates on how a direct enzyme-linked immunosorbent assay (ELISA) is used to specifically detect and quantify murine alpha-1 antitrypsin (AAT). As a direct ELISA, it lacks some sensitivity as compared to the "sandwich" ELISA method; however, it does reliably differentiate between samples with varying amounts of the mouse AAT protein. This protocol relies on the principle of adsorption to coat each well with sera proteins, whereas detection occurs specifically using a two-step antibody combination. This procedure effectively identifies and quantifies murine AAT from a wide variety of samples including mouse serum, cell culture medium, and cell or tissue lysate.

Rights and Permissions

Citation: Methods Mol Biol. 2017;1639:217-222. doi: 10.1007/978-1-4939-7163-3_21. Link to article on publisher's site

Related Resources

Link to Article in PubMed

Keywords

AAT, Alpha-1 antitrypsin, Direct ELISA, Enzyme-linked immunosorbent assay, Murine AAT

PubMed ID

28752461