Evaluation of protein phosphorylation during adipogenesis

Xi Li, Rong Zeng, Qi Qun Tang

Research output: Chapter in Book/Report/Conference proceedingChapter


Adipocyte differentiation is a complex process that involves the sequential expression of various adipocyte-specific genes controlled by signaling pathways and transcription factors for which phosphorylation plays a crucial regulatory role. CCAAT/enhancer-binding proteins and peroxisome proliferator-activated receptors are the most important transcriptional regulators in adipogenesis, and the functions of these proteins are regulated by various phosphorylation events. Because cultured 3T3-L1 preadipocytes are commonly used as a model for adipocyte differentiation, we used these cells for a proteomic analysis to identify kinases, phosphatases, and phosphosites that participate in adipogenesis. In addition to the phosphoproteomic analysis, we provide a detailed description of Western blotting, an in vitro phosphorylation assay, enzyme-linked immunosorbent assay, and phosphorylation site mutagenesis to fully characterize the phosphorylation of proteins and verify their roles in adipogenesis.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
Number of pages17
StatePublished - 2014
Externally publishedYes

Publication series

NameMethods in Enzymology
ISSN (Print)00766879
ISSN (Electronic)15577988


  • Adipogenesis
  • Phosphoproteomic analysis
  • Phosphorylation
  • Phosphorylation site mutagenesis
  • Sandwich ELISA for phosphoprotein

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology


Dive into the research topics of 'Evaluation of protein phosphorylation during adipogenesis'. Together they form a unique fingerprint.

Cite this