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Cold Spring Harb Protoc. 2018 Aug 13. doi: 10.1101/pdb.prot098376. [Epub ahead of print]

Mass Spectrometry-Based Absolute Quantification of Single Xenopus Embryo Proteomes.

Author information

1
Department of Molecular Biology, Faculty of Science, Radboud Institute for Molecular Life Sciences, Radboud University, Nijmegen 6500 HB, The Netherlands.
2
Genome Biology Unit, European Molecular Biology Laboratory, 69117 Heidelberg, Germany.
3
Department of Molecular Developmental Biology, Faculty of Science, Radboud Institute for Molecular Life Sciences, Radboud University, Nijmegen 6500 HB, The Netherlands.
4
Department of Molecular Biology, Faculty of Science, Radboud Institute for Molecular Life Sciences, Radboud University, Nijmegen 6500 HB, The Netherlands michiel.vermeulen@science.ru.nl.
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Contributed equally

Abstract

Early Xenopus development is characterized by a poor correlation between global mRNA and protein abundances due to maternal mRNA and protein loading. Therefore, proteome profiling is necessary to study gene expression dynamics during early Xenopus development. In contrast to mammals, single Xenopus eggs and embryos contain enough protein to allow identification and quantification of thousands of proteins using mass spectrometry-based proteomics. In addition to investigating developmental processes, single egg or blastomere proteomes can be used to study cell-to-cell variability at an unprecedented depth. In this protocol, we describe a mass spectrometry-based proteomics approach for the identification and absolute quantification of Xenopus laevis egg or embryo proteomes, including sample preparation, peptide fractionation and separation, and data analysis.

PMID:
30104410
DOI:
10.1101/pdb.prot098376

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