All reagents (unless otherwise stated) were purchased from Sigma-Aldrich Nümbrecht, Germany, and Sigma-Aldrich Saint Louis, MO, USA. Salivary glycoxidation and nitrosative stress assays were performed in duplicate samples. The absorbance/fluorescence was measured using Infinite M200 PRO Multimode Microplate Reader (Tecan Group Ltd., Männedorf, Switzerland). The results were standardized to 1 mg of total protein (TP).
TP content was analyzed using a commercial kit (Thermo Scientific PIERCE BCA Protein Assay; Rockford, IL, USA), according to the manufacturer's instructions. The salivary amylase activity was determined colorimetrically using 3,5-dinitrosalicylic acid (POCH, Gliwice, Poland) as a substrate reaction [40 (link)]. The absorbance was measured at 540 nm.
TP content was analyzed using a commercial kit (Thermo Scientific PIERCE BCA Protein Assay; Rockford, IL, USA), according to the manufacturer's instructions. The salivary amylase activity was determined colorimetrically using 3,5-dinitrosalicylic acid (POCH, Gliwice, Poland) as a substrate reaction [40 (link)]. The absorbance was measured at 540 nm.
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