Protocol for identification of proteins from deyolked zebrafish embryos.

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.