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Fig. 1. Gel-permeation chromatography of acute Cd-exposed (20 mg l–1, 2 days) N. diversicolor gut supernatant. Column: Hi- Load Superdex 75 Prep grade (Pharmacia); sample volume: 2 ml; flow rate: 1 ml min–1; elution buffer: 10 mmol l–1 Tris-HCl, pH 8.6. (A) The absorbance profile at 280 nm shows three main protein peaks, I, II and III. (B) Analysis of the Cd content (µg ml–1) of the chromatographic fractions shows that this metal is associated with components eluted in the void volume of the column (>70 kDa) likely to represent the hemoglobin (peak I in A) and components of the second main peak (peak II in A). (C) Enzyme-linked immunosorbent assay (ELISA) using the mAb anti-MPII revealed that peak II fractions contain the Cd-binding protein MPII.





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