Deliverables
Measurement methodology of metallothionein transcript level in the fish Lithognathus mormyrus, using real time PCR.
The result is an immuno-chemical method evaluating the level of cytochrome P4501A protein in fish liver. The utilized method is Enzyme-linked immuno-sorbent assay (ELISA), a general well established method for the evaluation of specific proteins in a mixture. The innovation of the present ELISA is by adapting this membrane protein to be used as ELISA target, by the production of heterologous modified cytochrome p4501A protein used as standard for quantification of the assay. The recombinant protein is soluble and can be bound to the walls of the ELISA in the first stage of the assay, enabling the utilization of a detergent in the second stage of the ELISA procedure to solubilise the native avaluated P4501A, detaching it from its membrane.
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