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Analysis of newly synthesized proteins by combined pulsed SILAC and click chemistry enrichment.
Analysis of newly synthesized proteins by combined pulsed SILAC and click chemistry enrichment. Experimental setup for the identification of proteins which are synthesized during depletion of UPF1. Pulse-labeling of cells with both the methionine analogue AHA and stable isotopes for 6 h allowed the enrichment of these proteins by click chemistry, on-bead trypsin digestion, fractionation of peptides and subsequent identification and quantification by LC-MS/MS (modified after (32)). Jana Sieber et al. Mol Cell Proteomics 2016;15: © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.
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