Protocol to quantify protein phosphorylation in mouse extended pluripotent stem cell-derived blastoids
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Updated: Aug 16, 2026

Quantitative Analysis of Protein Expression to Study Lineage Specification in Mouse Preimplantation Embryos
Published on: February 22, 2016
Tianxia Xiao1, Yuxin Luo2, Yang Yu3
1Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology and Key Laboratory of Assisted Reproduction, Ministry of Education, Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing 100191, China.
Abstract:
Protein phosphorylation is an important post-transcription modification regulating essential cellular activities. Here, we present a proteomics protocol to quantify protein phosphorylation in the extended pluripotent stem cell (EPSC)-derived blastoids from Mus musculus with a standardized phosphoproteome analysis workflow. We describe steps for culturing blastoids, preparing samples, enriching phosphopeptides, and fractionating peptides. We then detail procedures for acquiring and analyzing high-resolution liquid chromatography-mass spectrometry (LC-MS) data. This protocol provides a framework for reproducible analysis of phospho-proteomes of mouse blastoids. For complete details on the use and execution of this protocol, please refer to Min et al.1.

