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When melanin is overproduced or unevenly distributed, visible darkened areas can develop, presenting as hyperpigmentation or patchy tone
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Staining of formalin-fixed tissues requires heating tissue sections in 10mM Tris with 1mM EDTA, pH 9.0, for 45 min at 95 degrees C followed by cooling at RT for 20 minutes
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The samples were incubated on ice and centrifuged to separate the serum, which was then collected, transferred to clean tubes, and stored until further experimentation took place [2]
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