通过直接酶联免疫吸附测定法对小鼠α1-抗胰蛋白酶进行定量分析。

Quantification of Murine AAT by Direct ELISA.

作者信息

Cox Andrew, Mueller Christian

机构信息

Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA, USA.

Department of Pediatrics, University of Massachusetts Medical School, 368 Plantation St., AS6-2053, Worcester, MA, USA.

出版信息

Methods Mol Biol. 2017;1639:217-222. doi: 10.1007/978-1-4939-7163-3_21.

DOI:10.1007/978-1-4939-7163-3_21

PMID:28752461

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.

摘要

本章方法详细阐述了如何使用直接酶联免疫吸附测定（ELISA）来特异性检测和定量小鼠α-1抗胰蛋白酶（AAT）。作为一种直接ELISA，与“夹心”ELISA方法相比，它缺乏一定的灵敏度；然而，它确实能够可靠地区分含有不同量小鼠AAT蛋白的样本。该实验方案基于吸附原理，用血清蛋白包被每个孔，而检测则通过两步抗体组合特异性地进行。此程序可有效从多种样本中鉴定和定量小鼠AAT，这些样本包括小鼠血清、细胞培养基以及细胞或组织裂解物。