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The stirring continued for an additional 12 h at 50 C, after which the mixture was introduced to 10 mL of deionized water while being sonicated

Bio-Rad) using a 3-(N-morpholino) propane-sulfonic acid running buffer (Bio-Rad), and transferred onto polyvinylidene difluoride membranes (Bio-Rad)

Vitamin D: Update 2013: From rickets prophylaxis to general preventive healthcare