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Validation of the Architect 25-OH vitamin D assay.
Measurement of serum 25-OH-D was standardised; the hospital laboratory used the ARCHITEC T®25-OH Vitamin D assay (Abbott GmbH & Co KG, Wiesbaden-Delkenheim, Germany).
The LIAISON® 25-hydroxyvitamin D Assay (DiaSorin) uses chemiluminescent immunoassay technology.
We would like to thank Geeta T. Kumar for useful discussions about the vitamin D assay.
PLD activity was measured in 50 µg of cell extracts using the Amplex Red phospholipase D assay kit (Invitrogen Corporation, Carlsbad CA) according to the manufacturer's instructions.
The vitamin D assay was repeated by ZRT labs using the above procedure with 20 uL of whole blood or 10 ul serum for comparison.
We chose to follow up the Northern analysis above with the far more sensitive quantitative RT-PCR analysis and to combine this with an Actinomycin D assay in order to evaluate stability of the messages over time.
Activity of cathepsin B was assessed by monitoring hydrolysis of Z-LR-AMC (Enzo Life Sciences International), that of cathepsin D using the Cathepsin D Assay Kit (Sigma-Aldrich), and that of the 20S proteasome using the 20S Proteasome Assay Kit for Drug Discovery Enzo Life Sciences Internationall).
Blood was collected for 25-hydroxyvitamin D assay.
Spirometry was performed and blood taken for 25-hydroxyvitamin D assay.
Concern has been expressed about possible assay variability for the vitamin D assay in the NHANES (13).
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