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Standards (0.625–10 ng calf skin collagen I, Sigma Aldrich) and samples (100 µl) diluted with PBS (acid-soluble cell fraction, 1:10, and supernatants, 1:20), were loaded onto nitrocellulose membrane using a vacuum manifold to create protein dots

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Lack of this essential nutrient can cause several health problems, like fatigue, cardiac issues to everlasting neurological damages