A new technique for the isolation and fluorometric determination of polyadenosine diphosphate ribose in vivo

✍ Scribed by C. Niedergang; H. Okazaki; P. Mandel

• Publisher: Elsevier Science
• Year: 1978
• Tongue: English
• Weight: 501 KB
• Volume: 88
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(78)90394-9

✦ Synopsis

Polyadenosine 5'-diphosphate ribose (poly-ADPR) was isolated from animal tissue nuclei which had been extracted by the citric acid method. The isolation of poly-ADPR involved four steps: acidic precipitation of the homogenized nuclei; alkaline hydrolysis of the RNA and proteins; separation of poly-ADPR and DNA by cesium chloride equilibrium centrifugation; and chromatography on Sephadex G-10, enabling us to obtain poly-ADPR without DNA or RNA contamination and in water solution. The isolated poly-ADPR was broken down into its subunit, the ADPR, and then quantitatively determined by a fluorometric method modified for the determination of picomole quantities of ADPR.