Proteomics of early zebrafish embryos.
BACKGROUND
Zebrafish (D. rerio) has become a powerful and widely used model system for the analysis of vertebrate embryogenesis and organ development. While genetic methods are readily available in zebrafish, protocols for two dimensional (2D) gel electrophoresis and proteomics have yet to be developed.


RESULTS
As a prerequisite to carry out proteomic experiments with early zebrafish embryos, we developed a method to efficiently remove the yolk from large batches of embryos. This method enabled high resolution 2D gel electrophoresis and improved Western blotting considerably. Here, we provide detailed protocols for proteomics in zebrafish from sample preparation to mass spectrometry (MS), including a comparison of databases for MS identification of zebrafish proteins.


CONCLUSIONS
The provided protocols for proteomic analysis of early embryos enable research to be taken in novel directions in embryogenesis.
Link
Vinzenz
V
Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauer Str. 108, 01307 Dresden, Germany. link@mpi-cbg.de
Shevchenko
Andrej
A
Heisenberg
Carl-Philipp
CP
D000818:Animals; D015153:Blotting, Western; D030562:Databases, Protein; D004530:Egg Yolk / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D004625:Embryo, Nonmammalian / Q000737:chemistry / Q000378:metabolism; D013058:Mass Spectrometry; D040901:Proteomics / Q000379:methods*; D015027:Zebrafish / Q000196:embryology* / Q000378:metabolism; D029961:Zebrafish Proteins / Q000032:analysis* / Q000378:metabolism
D029961:Zebrafish Proteins