Protocol for high-throughput semi-automated label-free- or TMT-based phosphoproteome profiling.
STAR Protoc
; 4(3): 102536, 2023 Sep 15.
Article
em En
| MEDLINE
| ID: mdl-37659085
ABSTRACT
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.
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Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Fosfopeptídeos
/
Proteoma
Idioma:
En
Ano de publicação:
2023
Tipo de documento:
Article