Work

Protocol for identification of proteins from deyolked zebrafish embryos

Year: 2025

Type: article

Abstract: Zebrafish is a key model for studying vertebrate development and human diseases, but proteomic data during embryogenesis are limited due to interference from egg yolk proteins. Here, we present a protocol for the isolation, identification, and analysis of proteins from zebrafish embryos. We describe steps for dechorionation, deyolking, protein extraction, SDS-PAGE, and trypsin digestion. We then detail procedures for peptide separation using liquid chromatography-tandem mass spectrometry (LC-MS/MS), identification via ProteinPilot, and functional analysis with Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology. For complete details on the use and execution of this protocol, please refer to Purushothaman et al. 1 • Steps for deyolking to remove yolk proteins for proteomic analysis • Procedures for isolating proteins from yolk-rich embryos of amphibians and birds • Guidance on analyzing protein expression during early embryogenesis in vertebrates • Instructions for identifying and quantifying proteins using LC-MS/MS Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Zebrafish is a key model for studying vertebrate development and human diseases, but proteomic data during embryogenesis are limited due to interference from egg yolk proteins. Here, we present a protocol for the isolation, identification, and analysis of proteins from zebrafish embryos. We describe steps for dechorionation, deyolking, protein extraction, SDS-PAGE, and trypsin digestion. We then detail procedures for peptide separation using liquid chromatography-tandem mass spectrometry (LC-MS/MS), identification via ProteinPilot, and functional analysis with Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.103728