%0 Journal Article %1 kallerIdentificationMicroRNATargets2014 %A Kaller, Markus %A Oeljeklaus, Silke %A Warscheid, Bettina %A Hermeking, Heiko %C United States %D 2014 %J Methods in molecular biology (Clifton, N.J.) %K Acids/*chemistry,Analytic Amino Expression Gel,Gene High Labeling/*methods,Luciferases/genetics,MicroRNAs/*genetics,Proteolysis,Proteomics/*methods,Tandem Line Liquid,Electrophoresis Mass Methods,Antibodies/immunology,Blotting Neoplastic,Genetic Polyacrylamide Preparation Pressure Regulation Sample Spectrometry,to_read,Transfection,Trypsin/metabolism Tumor,Chromatography Vectors/genetics,Humans,Isotope Western,Cell %P 327--349 %R 10.1007/978-1-4939-1142-4_23 %T Identification of microRNA Targets by Pulsed SILAC. %V 1188 %X Pulsed stable isotope labeling by amino acids in cell culture (pulsed SILAC or pSILAC) allows to monitor and quantify the de novo synthesis of proteins in an unbiased fashion on a proteome-wide scale. The high applicability of this metabolic labeling technique has been demonstrated for the identification of posttranscriptional changes in gene expression on the proteome level, in particular those caused by microRNAs. The application of pSILAC allows the selective quantification of newly synthesized proteins and thus the detection of differences in protein translation. This is of particular interest in the case of microRNA-mediated regulations, which characteristically cause rather modest decreases in protein amounts that may be difficult to detect by other proteomic methods. Here, we describe a detailed protocol for using pSILAC to track miRNA-mediated changes in protein expression, using the p53-induced miR-34a microRNA as a prototypic example of microRNA-mediated regulations.

@article{kallerIdentificationMicroRNATargets2014, abstract = {Pulsed stable isotope labeling by amino acids in cell culture (pulsed SILAC or pSILAC) allows to monitor and quantify the de novo synthesis of proteins in an unbiased fashion on a proteome-wide scale. The high applicability of this metabolic labeling technique has been demonstrated for the identification of posttranscriptional changes in gene expression on the proteome level, in particular those caused by microRNAs. The application of pSILAC allows the selective quantification of newly synthesized proteins and thus the detection of differences in protein translation. This is of particular interest in the case of microRNA-mediated regulations, which characteristically cause rather modest decreases in protein amounts that may be difficult to detect by other proteomic methods. Here, we describe a detailed protocol for using pSILAC to track miRNA-mediated changes in protein expression, using the p53-induced miR-34a microRNA as a prototypic example of microRNA-mediated regulations.}, added-at = {2024-05-17T13:01:35.000+0200}, address = {United States}, author = {Kaller, Markus and Oeljeklaus, Silke and Warscheid, Bettina and Hermeking, Heiko}, biburl = {https://www.bibsonomy.org/bibtex/2b1f5215952a3a6dde44424aebcf71d03/warscheidlab}, doi = {10.1007/978-1-4939-1142-4_23}, interhash = {f8381f26d666bad86bff9c88618e0120}, intrahash = {b1f5215952a3a6dde44424aebcf71d03}, issn = {1940-6029 1064-3745}, journal = {Methods in molecular biology (Clifton, N.J.)}, keywords = {Acids/*chemistry,Analytic Amino Expression Gel,Gene High Labeling/*methods,Luciferases/genetics,MicroRNAs/*genetics,Proteolysis,Proteomics/*methods,Tandem Line Liquid,Electrophoresis Mass Methods,Antibodies/immunology,Blotting Neoplastic,Genetic Polyacrylamide Preparation Pressure Regulation Sample Spectrometry,to_read,Transfection,Trypsin/metabolism Tumor,Chromatography Vectors/genetics,Humans,Isotope Western,Cell}, langid = {english}, pages = {327--349}, pmid = {25059622}, timestamp = {2024-05-17T13:01:35.000+0200}, title = {Identification of {{microRNA}} Targets by Pulsed {{SILAC}}.}, volume = 1188, year = 2014 }