Sensitive quantitative analysis of disulfide bonds in polypeptides and proteins
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2021, LWTCitation Excerpt :Sample blanks contained 2.4 mL buffer with or without urea in 240 μL Ellman's reagent for the free or total SH group. We also characterized the S–S bonds content (Damodaran, 1985; Thannhauser, Konishi, & Scheraga, 1984). A 2-nitro-5-thiosulfobenzoate (NTSB) solution was prepared by adding 5, 5′-dithio-bis-2-nitrobenzoic acid (Sigma Aldrich, MO, USA) (0.1 g) into Na2CO3 buffer solution (10 mL, 1 M, pH 7.5) followed by continually flowing O2 into the solution at 38 °C until it turned orange.
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