title: Pulsed SILAC as a Approach for miRNA Targets Identification in Cell Culture.
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.
authors.affiliation: University of Cambridge Metabolic Research Laboratories and MRC Metabolic Diseases Unit, Wellcome Trust-MRC Institute of Metabolic Science, Addenbrooke's Hospital, Cambridge, UK.|Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil.
authors.forename: Daniella E
authors.initials: DE
authors.lastname: Duque-Guimarães
authors.affiliation: University of Cambridge Metabolic Research Laboratories and MRC Metabolic Diseases Unit, Wellcome Trust-MRC Institute of Metabolic Science, Addenbrooke's Hospital, Cambridge, UK.|Faculty of Medical Sciences, Department of Pharmacology, State University of Campinas, Campinas, Brazil.
authors.forename: Juliana
authors.initials: J
authors.lastname: de Almeida-Faria
authors.affiliation: University of Cambridge Metabolic Research Laboratories and MRC Metabolic Diseases Unit, Wellcome Trust-MRC Institute of Metabolic Science, Addenbrooke's Hospital, Cambridge, UK.|Food Research Center (FoRC) and Faculty of Pharmaceutical Sciences, University of São Paulo, São Paulo, Brazil.
authors.forename: Thomas Prates
authors.initials: TP
authors.lastname: Ong
authors.affiliation: University of Cambridge Metabolic Research Laboratories and MRC Metabolic Diseases Unit, Wellcome Trust-MRC Institute of Metabolic Science, Addenbrooke's Hospital, Cambridge, UK. seo10@cam.ac.uk.
authors.forename: Susan E
authors.initials: SE
authors.lastname: Ozanne
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D006801:Humans; D007553:Isotope Labeling; D035683:MicroRNAs / Q000235:genetics*; D014176:Protein Biosynthesis; D040901:Proteomics / Q000379:methods*; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism*; D012984:Software; D057188:Workflow
chemical_list: D000596:Amino Acids; D035683:MicroRNAs; D012333:RNA, Messenger
keywords: Cell culture; MicroRNAs; Proteomics; Pulsed stable isotope labeling by amino acids; Targets