Record number | 510288 |
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Title | In vivo identification of plant protein complexes using IP-MS/MS |
Author(s) | Wendrich, Jos R.; Boeren, Sjef; Möller, Barbara; Weijers, Dolf; Rybel, Bert de |
Source | In: Methods in Molecular Biology / Kleine-Vehn, Jürgen, Sauer, Michael, Humana Press Inc. (Methods in Molecular Biology ) - ISBN 9781493964673 - p. 147 - 158. |
DOI | http://dx.doi.org/10.1007/978-1-4939-6469-7_14 |
Department(s) |
Biochemistry EPS |
Publication type | Peer reviewed book chapter |
Publication year | 2017 |
Keyword(s) | Arabidopsis - GFP - Immunoprecipitation - Protein complex identification - Protein–protein interaction - Tandem mass spectrometry |
Abstract | Individual proteins often function as part of a protein complex. The identification of interacting proteins is therefore vital to understand the biological role and function of the studied protein. Here we describe a method for the in vivo identification of nuclear, cytoplasmic, and membrane-associated protein complexes from plant tissues using a strategy of immunoprecipitation followed by tandem mass spectrometry. By performing quantitative mass spectrometry measurements on biological triplicates, relative abundance of proteins in GFP-tagged complexes compared to background controls can be statistically evaluated to identify high-confidence interactors. We detail the entire workflow of this approach. |
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