Please use this identifier to cite or link to this item:
https://hdl.handle.net/2440/140433
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Type: | Journal article |
Title: | Endogenous protein interactomes resolved through immunoprecipitation-coupled quantitative proteomics in cell lines |
Author: | Kumar, R. Kamath, K.S. Carroll, L. Hoffmann, P. Gecz, J. Jolly, L.A. |
Citation: | STAR Protocols, 2022; 3(4):1-31 |
Publisher: | Elsevier BV |
Issue Date: | 2022 |
ISSN: | 2666-1667 2666-1667 |
Statement of Responsibility: | Raman Kumar, Karthik S. Kamath, Luke Carroll, Peter Hoffmann, Jozef Gecz, Lachlan A. Jolly |
Abstract: | Immunoprecipitation (IP) of endogenously expressed proteins is one of the most biologically relevant techniques to identify protein-protein interactions. We describe an adaptable IP protocol reliant on a specific antibody to the target protein. We detail a quantitative proteomics workflow for the unbiased identification of co-immunoprecipitating proteins, known collectively as an interactome. This includes protocols for the tryptic digestion, Tandem Mass Tag labeling and fractionation of peptides, and their identification and quantification using liquid chromatography-mass spectrometry including computational and statistical analysis. For complete details on the use and execution of this protocol, please refer to Johnson et al. (2020). |
Keywords: | Cell biology Mass spectrometry Protein expression and purification Proteomics |
Rights: | © 2022 The Author(s). This article is available under the Creative Commons CC-BY-NC-ND license and permits non-commercial use of the work as published, without adaptation or alteration provided the work is fully attributed. |
DOI: | 10.1016/j.xpro.2022.101693 |
Grant ID: | http://purl.org/au-research/grants/nhmrc/1155224 http://purl.org/au-research/grants/arc/170103090 |
Published version: | http://dx.doi.org/10.1016/j.xpro.2022.101693 |
Appears in Collections: | Research Outputs |
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File | Description | Size | Format | |
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hdl_140433.pdf | Published version | 4.65 MB | Adobe PDF | View/Open |
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