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QuaNCAT: quantitating proteome dynamics in primary cells.

Nature methods (2013-03-12)
Andrew J M Howden, Vincent Geoghegan, Kristin Katsch, Georgios Efstathiou, Bhaskar Bhushan, Omar Boutureira, Benjamin Thomas, David C Trudgian, Benedikt M Kessler, Daniela C Dieterich, Benjamin G Davis, Oreste Acuto
ABSTRACT

Here we demonstrate quantitation of stimuli-induced proteome dynamics in primary cells by combining the power of bio-orthogonal noncanonical amino acid tagging (BONCAT) and stable-isotope labeling of amino acids in cell culture (SILAC). In conjunction with nanoscale liquid chromatography-tandem mass spectrometry (nanoLC-MS/MS), quantitative noncanonical amino acid tagging (QuaNCAT) allowed us to monitor the early expression changes of >600 proteins in primary resting T cells subjected to activation stimuli.

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Ionomycin from Streptomyces conglobatus, ≥98% (HPLC)