A highly sensitive method for large-scale measurements of 1,25-dihydroxyvitamin D

Analytical Biochemistry
N C ArbourH F DeLuca

Abstract

A quantitative method for measuring 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) was developed utilizing a luciferase reporter gene under the control of the highly inducible 25-hydroxyvitamin D3 24-hydroxylase promoter in a stably transfected cell line. Transient transfections with constructs containing the 24-hydroxylase gene promoter 5' to a luciferase reporter were first performed in cell lines with high levels of vitamin D receptor, i.e., the rat osteosarcoma (ROS 17/2.8) and human breast cancer (T-47D) cell lines. ROS 17/2.8 cells, stably transfected with the plasmid, gave a 60-fold stimulation with 10(-10) M 1,25-(OH)2D3. A standard curve was constructed showing a large range of response to 1,25-(OH)2D3 (1 pg to 1 ng). The assay was adapted to microtiter plates, which permits a large number of samples to be assayed simultaneously. Other metabolites of vitamin D and analogs such as 25-hydroxyvitamin D3, 24,25-dihydroxyvitamin D3, and 1 alpha-hydroxyvitamin D3 have negligible effects on the detection of 1,25-(OH)2D3, thus eliminating the need for purification of sample. The sensitivity of the method permitted the use of 100 microliters of serum with excellent results. Comparison of this method with a commercially available assay...Continue Reading

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Citations

Sep 11, 2013·Proceedings of the National Academy of Sciences of the United States of America·Jinge G ZhuHector F Deluca
May 1, 2004·Proceedings of the National Academy of Sciences of the United States of America·Lori A PlumHector F DeLuca
Nov 3, 1998·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·C GrossD Feldman
Jun 15, 2011·Molecular and Cellular Endocrinology·Frank BienaiméJean Claude Souberbielle
Mar 11, 2010·The Journal of Steroid Biochemistry and Molecular Biology·Agnieszka GlebockaHector F DeLuca
Mar 30, 2010·The Journal of Steroid Biochemistry and Molecular Biology·Izabela SibilskaHector F Deluca
Feb 16, 2010·The Journal of Steroid Biochemistry and Molecular Biology·Katarzyna SokolowskaHector F DeLuca
Oct 1, 2008·The Journal of Steroid Biochemistry and Molecular Biology·Alanna Roff, Robin Taylor Wilson
Aug 17, 1999·Biochemical Pharmacology·L H Naylor
Jan 1, 2013·The Journal of Steroid Biochemistry and Molecular Biology·Izabela SibilskaHector F DeLuca
Oct 16, 2012·The Journal of Steroid Biochemistry and Molecular Biology·Agnieszka GlebockaHector F Deluca
Dec 29, 2012·The Journal of Steroid Biochemistry and Molecular Biology·Urszula KuleszaRafal R Sicinski
Nov 3, 2012·The Journal of Steroid Biochemistry and Molecular Biology·Katarzyna SokolowskaHector F DeLuca
Jan 16, 2007·The Journal of Steroid Biochemistry and Molecular Biology·Katarzyna Plonska-OcypaHector F DeLuca
Mar 13, 2017·The Journal of Steroid Biochemistry and Molecular Biology·Marcin SzybinskiHector F DeLuca
Jul 1, 2004·The Journal of Steroid Biochemistry and Molecular Biology·Yuko OdaDaniel D Bikle
Sep 9, 2006·Journal of Cellular Biochemistry·Anna KlopotJames C Fleet
Oct 6, 2005·The Journal of Biological Chemistry·Rubina YasminNoa Noy
Nov 27, 2014·American Journal of Physiology. Endocrinology and Metabolism·Yongji WangHector F DeLuca
Jun 26, 1999·The Journal of Biological Chemistry·A Racz, J Barsony
Oct 23, 2010·Archives of Biochemistry and Biophysics·Hector F DelucaLori A Plum
Jun 11, 2002·Archives of Biochemistry and Biophysics·Matthew J Beckman, Hector F DeLuca
Nov 15, 2014·The Journal of Organic Chemistry·Katarzyna SokolowskaRafal R Sicinski

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