Figure 4 - Size exclusion in HPLC of trypsinogen urea denaturation. The column (Shodex - Protein KW 803 (0.8 x 30 cm), Showa Denko & Shoko) was loaded with 50 µg of protein per run and equilibrated with 100 mM MES, pH 7.0, 20 mM CaCl2 and 81 mM NaNO3, pH 7.0, containing urea at different concentrations. Trypsinogen concentration was 1 mM in each sample. The flow rate was 0.75 ml/min and elution was monitored by absorbance at 280 nm. Profiles present the retention times in minutes for each urea concentration. The Stokes radii were calculated according to Uversky (21).