Figure 1 | Scientific Reports

Figure 1

From: Proteomics of thyroid tumours provides new insights into their molecular composition and changes associated with malignancy

Figure 1

Overview of sample preparation and analysis.

(a) Fresh frozen tissues from histologically normal (N), FA, FTC and PTC samples were lysed and subjected to protein reduction, alkylation and tryptic digestion; (b) generation of spectral library for SWATH-MS was achieved by combination of independent IDA-MS/MS experiments from six unfractionated and eight SAX-fractionated samples, which allowed downstream data processing including peptide identification and quantitative analysis; (c) iTRAQ-MS and MRM-HR were employed as validation techniques; the former was carried out by 4-plex labelling of FA, FTC, PTC and masterpool samples subsequently mixed, fractionated by SCX chromatography and loaded onto the LC-MS system. MRM-HR of selected proteins/peptides was performed targeting specific precursor m/z values, collecting high-resolution MS/MS spectra and extracting quantitative information from selected fragment ions.

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