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After TMB substrate solution is added, only those wells that contain Mouse ACTH, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color

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Expression of these proteins [Gsta1-2-3, glutathione- S -transferase theta 1a (Gstt1a), Gstz1-2-3, Gstm3, and Gsto1] increased steadily from 72 hpf to 168 hpf, while in adults only Gsta1-2-3, Gstt1a, and Gst1, 2, 3 proteins were found to be liver enriched suggesting these might be the main Gsts of phase II detoxification in zebrafish liver maturation [24]