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{"title":"Tandem Affinity Purification and Mass Spectrometry (TAP-MS) for the Analysis of Protein Complexes","authors":"Guillaume Adelmant, Brijesh K. Garg, Maria Tavares, Joseph D. Card, Jarrod A. Marto","doi":"10.1002/cpps.84","DOIUrl":null,"url":null,"abstract":"<p>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.</p>","PeriodicalId":10866,"journal":{"name":"Current Protocols in Protein Science","volume":"96 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2019-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpps.84","citationCount":"16","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Protein Science","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpps.84","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
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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.
串联亲和纯化和质谱(TAP-MS)分析蛋白质复合物
亲和纯化后的质谱法已成为鉴定从生理细胞环境分离的生化复合物结合伙伴的首选技术。在本报告中，我们详细介绍了我们的串联亲和纯化(TAP)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，以及实现自动化版本TAP程序的步骤。©2019 by John Wiley &儿子,Inc。
本文章由计算机程序翻译，如有差异，请以英文原文为准。
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