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Updated: May 16, 2025
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Published on: August 31, 2016
Protocol for identification of proteins from deyolked zebrafish embryos
Kathiresan Purushothaman1, Saraphina Dianne Tneo Rwei Qing2, Yong Xin Ee3
1Genomics Group, Faculty of Biosciences and Aquaculture, Nord University, 8049 Bodø, Norway; School of Applied Science, Republic Polytechnic, 9 Woodlands Avenue 9, Singapore 738964, Singapore; Department of Preclinical Sciences and Pathology, Faculty of Veterinary Medicine, Norwegian University of Life Sciences, Ås, Norway; Tropical Futures Institute, James Cook University Singapore, 149 Sims Drive, Singapore 387380, Singapore.
None:
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.
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