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用于蛋白质复合物分析的串联亲和纯化与质谱联用技术（TAP-MS）

Tandem Affinity Purification and Mass Spectrometry (TAP-MS) for the Analysis of Protein Complexes.

作者信息

Adelmant Guillaume, Garg Brijesh K, Tavares Maria, Card Joseph D, Marto Jarrod A

机构信息

Department of Cancer Biology and Blais Proteomics Center, Dana-Farber Cancer Institute, Boston, Massachusetts.

Department of Pathology, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts.

出版信息

Curr Protoc Protein Sci. 2019 Jun;96(1):e84. doi: 10.1002/cpps.84. Epub 2019 Feb 1.

DOI:10.1002/cpps.84
PMID:30706993
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6579647/
Abstract

Affinity purification followed by mass spectrometry has become the technique of choice to identify binding partners in biochemical complexes isolated from a physiologic cellular context. In this report we detail our protocol for tandem affinity purification (TAP) primarily based on the use of the FLAG and HA peptide epitopes, with a particular emphasis on factors affecting yield and specificity, as well as steps to implement an automated version of the TAP procedure. © 2019 by John Wiley & Sons, Inc.

摘要

亲和纯化后进行质谱分析已成为从生理细胞环境中分离出的生化复合物中鉴定结合伴侣的首选技术。在本报告中，我们详细介绍了串联亲和纯化（TAP）方案，该方案主要基于FLAG和HA肽表位的使用，特别强调了影响产量和特异性的因素，以及实施TAP程序自动化版本的步骤。© 2019约翰威立父子公司版权所有