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A total of 20 g protein was separated using 10% or 12.5% sodium dodecyl sulphate polyacrylamide gel electrophoresis and blotted onto polyvinylidene fluoride membranes, which were then blocked with 5% non-fat milk diluted in Tris-buffered saline containing 0.05% Tween 20 for 1 h at room temperature

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Also adding in Vitamin-D drops particularly in winter can assist to help regulate melatonin, calcium and improve sleep

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