The study of direct ELISA and competitive ELISA for rabbit metallothionein: Correlation of induction with zinc

Abstract

On the basis of the purification of metallothionein (MT) from human fetal liver, the purified polyclonal antibody of rabbit anti-human MT-1 was prepared. Direct enzyme-linked immunosorbent assay (ELISA) for MT in rabbit tissues was established. Direct ELISA is a direct binding assay. Its sensitivity is 0.4 ng (4 ng/ml). MT contents of control rabbit liver and kidney are 240 μg/g and 200 μg/g wet weight tissue, respectively. MT contents of high induction rabbit liver and kidney are 3130 μg/g and 3570 μg/g wet weight tissue, respectively. Competitive ELISA for MT in erythrocyte lysate was determined. Coating concentration is 20 ng/ml; antibody dilution is 1:2000; competitive MT concentrations are 40-2500 ng/ml. Its sensitivity extends to 2 ng (40 ng/ml). MT concentration in the control rabbit erythrocyte lysate is 10.52 μg/g total lysate protein; it is 43.16 μg/g total lysate protein in the low induction group, and 94.93 mg/g total lysate protein in the high induction group.