A&B. TRAMP mice harbouring the tx mutation (tx/TRAMP) display elevated copper in their liver and serum. Inductively coupled plasma mass spectrometry (ICP-MS) was used to determine copper concentrations in the liver and serum of 22-week old wild type, tx, TRAMP and tx/TRAMP mice (n=5-20 for each strain). Results represent mean ± STDEV (whisker plot) and are shown as either μg/g wet weight for tissue or μM for serum. C. Copper incorporation into serum ceruloplasmin is perturbed in TRAMP mice harbouring the tx mutation (tx/TRAMP). Serum ceruloplasmin oxidase activity (copper-dependent) was measured from 22-week old wild type (n=10), tx (n=11), TRAMP (n=17) and tx/TRAMP (n=10) mice using the o-dianisidine dihydrochloride based assay. Results are expressed as unit/litre (U/L) and presented as mean ± STDEV (bar). D-H. TRAMP mice harbouring the tx mutation (tx/TRAMP) display elevated copper in extrahepatic tissues. ICP-MS was used to determine copper concentrations in brain, kidney, spleen, lung and prostate lobes [anterior prostate (AP), dorsolateral prostate (DLP) and ventral prostate (VP)] of 22-week old wild type, tx, TRAMP and tx/TRAMP mice (n=4-20 for each strain). Results represent mean ± STDEV (whisker plot) and are shown as μg/g wet weight. I. Mouse prostate has no detectable level of Atp7b expression. (i) Real-time PCR quantification of Atp7b mRNA levels in liver (LIV), kidney (KID) and prostate (PRO) of 22-week old wild type mice (n=3). The level of Atp7b mRNA is compared against the liver. (ii) Western blot analysis of Atp7b expression in liver (LIV), kidney (KID) and prostate (PRO) of 22-week old wild type mice (50 μg protein). The WND4B antibody detected Atp7b in the liver and kidney at ~170kDa. β-actin was detected as a loading control. (*p < 0.05; **p < 0.01; ***p < 0.001).