The primary synthesis of functional vitamin D begins in the skin, where a cholesterol by-product acted upon by UV light forms vitamin D3. Vitamin D3 is carried through the bloodstream, attached to the vitamin D binding protein, to the liver where it is converted to 25-hydroxy vitamin D3 (25(OH)D3). This product is then transported to the kidney where it is further converted to 1,25-dihydroxycholecalciferol (1,25(OH)2D3), the biologically active form. Active 1,25(OH)2D3 is very short-lived and is rapidly metabolised to its deactivated forms.
Cayman Chemical’s Vitamin D ELISA Kit is a competitive assay that can be used for vitamin D quantification in plasma or serum. Because of the short half-life of the biologically active 1,25(OH)D form, the vitamin D assay primarily detects the more metabolically stable forms, 25(OH)D3 and 25(OH)D2. Detection of these forms requires that they be displaced from the vitamin D binding protein prior to measurement. A simple, acetone-based purification protocol is included as the initial procedure of the vitamin D assay kit for this purpose.
With an intubation time of 18 h and a development time of 60–90 min, the kit can measure vitamin D levels down to 0.5 ng/mL. It can assay 24 samples in triplicate or 36 samples in duplicate.
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