Protocol for identification of proteins from deyolked zebrafish embryos
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.<sup>1</sup>.
Authors
- Qingsong Lin (ORCID: https://orcid.org/0000-0001-9117-8514)
- Shubha Vij (ORCID: https://orcid.org/0000-0003-1159-2518)
- Igor Babiak (ORCID: https://orcid.org/0000-0002-1462-2690)
- Saraphina Dianne Tneo Rwei Qing
- Kathiresan Purushothaman
- Yong Xin Ee
- Nithiyakala Ananthakrishnan
- Shanmugam Harshiktha
Institutions
- Madras Medical College (IN)
- National University of Singapore (SG)
- Republic Polytechnic (SG)
- Nord University (NO)
- ASA College (US)
- James Cook University Singapore (SG)
- Université Bourgogne Franche-Comté (FR)
- Norwegian University of Life Sciences (NO)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-04-07
- DOI
- https://doi.org/10.1016/j.xpro.2025.103728
- Citations
- 1
- Primary Topic
- Aquaculture disease management and microbiota
- Type
- article
- Field-Weighted Citation Impact
- 0.80