Pierce trypsin protease
Pierce trypsin protease is a proteolytic enzyme used for the digestion and cleavage of proteins. It functions by catalyzing the hydrolysis of peptide bonds, primarily at the carboxyl side of lysine and arginine residues.
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Quantitative Proteomic Sample Preparation
C18 columns, Pierce trypsin protease, lysis buffer, Pierce protease inhibitor tablets and lysyl-endopeptidase LysC were purchased from Thermo Scientific (Rockford, IL, USA). Bovine serum albumin and Bradford’s reagent were purchased from Sigma-Aldrich (St. Louis, MO, USA).
Quantitative Protein Profiling by TMT
For quantitative proteomics, each digested protein sample was labelled with an appropriate TMT2plex Isobaric Label Reagent (Thermo Scientific, MA), according to the protocol supplied by the manufacturer. Briefly, the control and treated peptide samples were labelled with reagents TMT-126 and TMT-127, respectively. To quench the TMT reaction, 8 μL of 5% hydroxylamine was added to each sample and incubated for 15 min at room temperature. All the labelled peptides were combined in equal amounts and dried using a speed vacuum system.
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