A nonradioactive assay for poly(a)-specific ribonuclease activity by methylene blue colorimetry

Protein Pept Lett. 2006;13(2):125-8. doi: 10.2174/092986606775101580.

Abstract

A simple nonradioactive assay, which was based on the specific shift of the absorbance maximum of methylene blue induced by its intercalation into poly(A) molecules, was developed for poly(A)-specific ribonuclease (PARN). A good linear relationship was found between the absorbance at 662 nm and the poly(A) concentration. The assay conditions, including the concentration of methylene blue, the incubation temperature and time, and the poly(A) concentration were evaluated and optimized.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Colorimetry / methods*
  • Diagnostic Techniques, Radioisotope
  • Exoribonucleases / metabolism*
  • Humans
  • Methylene Blue / analysis*
  • Poly A / metabolism*
  • Spectrum Analysis
  • Substrate Specificity
  • Temperature

Substances

  • Poly A
  • Exoribonucleases
  • poly(A)-specific ribonuclease
  • Methylene Blue