The samples were stored at −80 °C until preparation was initiated. By the beginning of the preparation, the samples were thawed and Schirmer strips incubated for 20 min in lysis buffer (5% SDS, 50 mM triethylammonium bicarbonate [TEAB]). Samples were prepared according to the S-Trap™ Micro spin column digestion protocol from ProtiFi (ProtiFi, Huntington, NY, USA) as previously described [8 (link)].
The reduction in disulphide bonds, alkylation of cysteines, and digestion in S-Trap micro columns were performed as described in a recent article [8 (link)]. The elution of peptides, recovery of hydrophobic peptides, and measurement of peptide concentration were performed as previously described [8 (link),30 (link)]. Each sample was dried in a vacuum centrifuge and stored at −80 °C until further use.
The reduction in disulphide bonds, alkylation of cysteines, and digestion in S-Trap micro columns were performed as described in a recent article [8 (link)]. The elution of peptides, recovery of hydrophobic peptides, and measurement of peptide concentration were performed as previously described [8 (link),30 (link)]. Each sample was dried in a vacuum centrifuge and stored at −80 °C until further use.
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