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Supernatant was collected (490 l of cytosolic fraction) while cellular membranes (P100) were washed once with 1X PBS and re-suspended in 50 l Tris (10 mM, pH 7.4)

To investigate alterations to MT3 in the human ALS and MS spinal cord, and how these relate to levels of copper and zinc, we applied parallel mass spectrometry approaches in conjunction with immunohistochemistry (Fig