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Pulsed SILAC as a Approach for miRNA Targets Identification in Cell Culture
Daniella E Duque-Guimarães1,2, Juliana de Almeida-Faria1,3, Thomas Prates Ong1,4
1University of Cambridge Metabolic Research Laboratories and MRC Metabolic Diseases Unit, Wellcome Trust-MRC Institute of Metabolic Science, Addenbrooke's Hospital, Cambridge, UK.
Methods in Molecular Biology (Clifton, N.J.)
|November 30, 2016
Abstract:
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.