PMID: 3766947Aug 1, 1986Paper

A new highly selective physicochemical assay to measure NAD+ in intact cells

Analytical Biochemistry
R Alvarez-GonzalezF R Althaus

Abstract

A simple, fast, and highly specific chromatographic method for measuring the content of NAD+ in intact cells has been developed. This procedure involves the separation of NAD+ from the bulk of acid-soluble nucleosides, nucleotides, and other pyridine containing molecules by affinity chromatography on dihydroxyboronyl-Bio-Rex. The boronate purified preparations were utilized for the quantification of NAD+ by strong anion exchange high-pressure liquid chromatography under isocratic conditions using a low salt buffer system. The overall recovery of the method exceeded 80%. This new method was applied to determine the extent of NAD+ consumption in intact hepatocytes following treatment with two different DNA damaging agents. A major advantage of this method is that it allows for the simultaneous determination of poly(ADP-ribose) in the acid-insoluble fraction of the same sample.

References

Aug 1, 1976·Archives of Biochemistry and Biophysics·E L Jacobson, M K Jacobson
Dec 13, 1979·Nature·H Juarez-SalinasM K Jacobson
Aug 15, 1985·The Biochemical Journal·J C Gaal, C K Pearson
Jan 1, 1985·Annual Review of Biochemistry·K Ueda, O Hayaishi
Jan 1, 1985·Current Topics in Cellular Regulation·N O Kaplan
Dec 17, 1985·Biochemistry·D M PayneM K Jacobson
Mar 1, 1980·Analytical Biochemistry·C Labarca, K Paigen
Feb 7, 1980·Nature·B W DurkaczS Shall
Nov 1, 1983·Analytical Biochemistry·R Alvarez-GonzalezM K Jacobson

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Citations

Oct 6, 2000·Archives of Biochemistry and Biophysics·L AtorinoP Quesada
Dec 15, 1994·Molecular & General Genetics : MGG·M A Collinge, F R Althaus
Sep 1, 1989·Mutation Research·R Alvarez-Gonzalez, F R Althaus
Mar 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·P LoetscherF R Althaus

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