Quantitative proteomics : TMT-based quantitation of proteins

Quantification of proteins using isobaric labeling (tandem mass tag or TMT) starts with the reduction of disulfide bonds in proteins with Dithiothreitol (DTT). Alkylation with iodoacetamide (IAA) after cystine reduction results in the covalent addition of a carbamidomethyl group that prevents the formation of disulfide bonds. Then, overnight digestion of the proteins using trypsin or trypsin/LyC
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Quantitative proteomics: label-free quantitation of proteins

Liquid chromatography (LC) coupled with mass spectrometry (MS) has been widely used for protein expression quantification. Protein quantification by tandem-MS (MS/MS) uses integrated peak intensity from the parent-ion mass (MS1) or features from fragment-ions (MS2). Label-free quantification (LFQ) may be based on precursor ion intensity (peak areas or peak heights) or on spectral counting. Here, the
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