An alkaline phosphatase-linked assay for ribonucleases

✍ Scribed by Robert Stern; Joseph Wilczek

• Publisher: Elsevier Science
• Year: 1973
• Tongue: English
• Weight: 588 KB
• Volume: 54
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(73)90371-0

✦ Synopsis

A convenient and rapid assay for ribonucleases has been developed using commerical unlabeled materials. This assay detected less than 1 ng of RXase A. The assay was also applied to RNase T, and micrococcal nuclease. The phosphate end groups generated at the cleavage sites of the RNA substrate were measured by incubating with cscess alkaline phosphatase and determining the phosphate released. Initial reaction rates were measured and accurate units of activity established, which is not possible with most R?;ase assays. Commercial preparariona of alkaline phosphatase from E. coli are contaminated with RKase. A procedure was described for removal of RNase from the alkaline phosphatase preparations.