After normalizing the extracted ion chromatogram with respect to the internal standard (vitamin D3), the signal enhancement was consistently 2-3 orders of magnitude greater than the negative control using both peak height and peak area as the experimental criteria. This assay reliably demonstrates binding of the VDR with its endogenous ligand calcitriol, and will effectively serve as the positive control for screening extracts against this biologically relevant target. The development of this assay complements the myriad of PUF MS assays already in use by this lab such as screens for ligands of cyclooxygenase-2 and the estrogen receptors which are currently being applied to the discovery of natural chemoprevention agents from marine sediments bacteria and botanical extracts.
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