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To study and quantify the oxidation of thiols within the multilayers, 1 ¦Ìm silica particles coated with five bilayers of PMASH/PVPON were used. In
reduced form, the multilayers are unstable at pH 7 and rapidly disassemble when placed into a 10 mM solution of Ellman¡¯s reagent in 1 M
phosphate buffer, pH 7.5. The resulting solution has a characteristic absorption at 412 nm (Figure 2, spectrum 1), which is used to quantify the
number of thiol groups by using a calibration curve obtained with standard solutions of glutathione. The addition of CaT to the particles with
PMASH/PVPON multilayers at different pH (4¨C6) results in a decreased number of thiol groups, reflecting their oxidation into disulfide linkages. We found that the addition of 2 mole equivalents of CaT oxidized 90% of thiols within 1 min (Figure 2, spectrum 2), and a moderate excess of CaT (2
mM solution) converts the thiols into disulfides nearly quantitatively (Figure 2, spectrum 3).
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