Protocol for high-throughput semi-automated label-free- or TMT-based phosphoproteome profiling
Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
Dokumenter
- Fulltext
Forlagets udgivne version, 8,28 MB, PDF-dokument
Tandem mass tags data-dependent acquisition (TMT-DDA) as well as data-independent acquisition-based label-free quantification (LFQ-DIA) have become the leading workflows to achieve deep proteome and phosphoproteome profiles. We present a modular pipeline for TMT-DDA and LFQ-DIA that integrates steps to perform scalable phosphoproteome profiling, including protein lysate extraction, clean-up, digestion, phosphopeptide enrichment, and TMT-labeling. We also detail peptide and/or phosphopeptide fractionation and pre-mass spectrometry desalting and provide researchers guidance on choosing the best workflow based on sample number and input. For complete details on the use and execution of this protocol, please refer to Koenig et al.1 and Martínez-Val et al.2
Originalsprog | Engelsk |
---|---|
Artikelnummer | 102536 |
Tidsskrift | STAR Protocols |
Vol/bind | 4 |
Udgave nummer | 3 |
Antal sider | 30 |
ISSN | 2666-1667 |
DOI | |
Status | Udgivet - 2023 |
Bibliografisk note
Funding Information:
Work at the Novo Nordisk Foundation Center for Protein Research (CPR) is funded in part by a generous donation from the Novo Nordisk Foundation ( NNF14CC0001 ). This work has also been funded as part of the EPIC-XS project under the grant agreement no. 823839 funded by the Horizon 2020 program of the European Union and supported by the European Research Council through ERC-Synergy grant 810057 -HighResCells. C.K. is supported by the Marie Skłodowska Curie European Training Network “PUSHH” (grant no. 861389 ).
Publisher Copyright:
© 2023 The Authors
Antal downloads er baseret på statistik fra Google Scholar og www.ku.dk
ID: 367712319