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Afterward, the samples were washed 3 times in PBS-T [0.2% (v/v) Tween 20 in PBS] and incubated with secondary antibodies: Cy3-conjugated donkey anti-rat secondary antibody (ImmunoResearch Europe, 712-166-153

Avoid EDTA, DTPA, or other strong chelators in buffer systems as they will strip copper from the complex

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In research models where caloric intake is unrestricted, this is typically well-tolerated

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