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Quantification of Murine AAT by Direct ELISA
Andrew Cox1, Christian Mueller2,3
1Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|July 29, 2017
Summary
This study details a direct enzyme-linked immunosorbent assay (ELISA) for detecting mouse alpha-1 antitrypsin (AAT). The method reliably quantifies AAT in various biological samples, despite lower sensitivity than sandwich ELISA.
Area of Science:
- Biochemistry
- Immunology
- Assay Development
Background:
- Alpha-1 antitrypsin (AAT) is a crucial proteinase inhibitor.
- Accurate quantification of murine AAT is essential for research.
- Existing methods may have limitations in sensitivity or sample compatibility.
Purpose of the Study:
- To describe a direct enzyme-linked immunosorbent assay (ELISA) protocol.
- To enable specific detection and quantification of murine alpha-1 antitrypsin (AAT).
- To provide a reliable method for AAT measurement in diverse biological matrices.
Main Methods:
- Utilizes a direct ELISA format.
- Employs adsorption to coat wells with sera proteins.
- Features a two-step antibody combination for specific detection.
Main Results:
- Successfully detects and quantifies murine AAT.
- Demonstrates reliable differentiation between samples with varying AAT concentrations.
- Applicable to mouse serum, cell culture medium, and cell/tissue lysates.
Conclusions:
- The direct ELISA is a viable method for murine AAT quantification.
- This protocol offers a straightforward approach for AAT measurement.
- The method is robust across multiple sample types.
