2. Academic Validation
  3. Protocol to quantify protein phosphorylation in mouse extended pluripotent stem cell-derived blastoids

Protocol to quantify protein phosphorylation in mouse extended pluripotent stem cell-derived blastoids

  • STAR Protoc. 2025 Nov 3;6(4):104166. doi: 10.1016/j.xpro.2025.104166.
Tianxia Xiao 1,  Yuxin Luo 2,  Yang Yu 3
Affiliations

Affiliations

  • 1 Beijing 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.
  • 2 Stem Cell Research Center, Peking University Third Hospital, Beijing 100191, China.
  • 3 Beijing 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; Stem Cell Research Center, Peking University Third Hospital, Beijing 100191, China. Electronic address: [email protected].
PMID: 41191480 DOI: 10.1016/j.xpro.2025.104166
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.

Keywords

Developmental biology; Proteomics; Stem Cells; Systems biology.

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