Purification, characterization and ELISA of metallothionein from red scorponifish, Sebastiscus marmoratus

Metallothionein(MT) is a low molecular weight (MW)and heatstable metal-binding prorein.It has been proved that the synthesis of MT in marine animals can be induced by various heavy metals at genetic level.Consequently, high level of MT can be used as sign of heavy metal pollution.
In this article, red scorponifish MT was purified by Sephadex G-75 gel filtration and DEAE-Sepharose CL-6B ion exchange chromatography.The results of amino acids composition analysis showed that the protein contains 35%cysteine (Cys),but no aromatic residues.MW was ahout 6.7 kD, and pI is 4.0 and 4.2,respectively.The antibody was raised in white rabbits by injection by MT conjugated with BSA, and then purified on Protein A beads and labeled with horseradish peroxidase.Quantification of MT by ELISA indicated that the concentration of MT from induced and noninduced fish liver extraction were 450 and 20 μg/cm³,respectively.