Pulsed SILAC as a Approach for miRNA Targets Identification in Cell Culture.

Daniella E Duque-Guimarães, Juliana de Almeida-Faria, Thomas Prates Ong, Susan E Ozanne

OAI: oai:www.repository.cam.ac.uk:1810/278050 • DOI: 10.17863/CAM.25390

Pulsed stable isotope labeling by amino acids in cell culture (pSILAC) comprises a variation of the classical SILAC proteomic methodology that enables the identification of short-term proteomic responses such as those elicited by micro RNAs (miRNAs). Here, we describe a detailed pSILAC protocol for global identification and quantification of protein translation alterations induced by a miRNA using 3T3-L1 pre-adipocytes as a model system.

Cell culture MicroRNAs Proteomics Pulsed stable isotope labeling by amino acids Targets Amino Acids Cell Culture Techniques Cells Cultured Gene Expression Regulation Humans Isotope Labeling MicroRNAs Protein Biosynthesis Proteomics RNA Interference RNA Messenger Software Workflow