Assay of (2′-5′)-Oligo(A) Synthesized in Vitro and the Analysis of Naturally Occurring (2′-5′)-Oligo(A) from Intact Cells

B. R.G. Williams, R. E. Brown, C. S. Gilbert, R. R. Golgher, D. H. Wreschner, W. K. Roberts, R. H. Silverman, Ian M. Kerr

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Researchpeer-review

17 Citations (Scopus)

Abstract

This chapter presents methods used for radioactive ATP incorporation and biological assays. The wide distribution of the 2-5A synthetase in a variety of cells and tissues and its variation with growth and hormone status have emphasized the wider possible significance of the 2-5A system and the desirability of assay methods for the routine estimation of naturally occurring preformed 2-5A. Accordingly, the methods that have been used successfully in the initial experiments on the recovery of 2-5A from intact cells are also presented. The 2-5A is effective at nanomolar concentrations in cell-free systems. The detection of 2-5A at concentrations lower than this has been complicated by the nonspecific inhibitory effects of the crude extracts in the biological assay employed, the sensitivity of the assays, and the amounts of starting material readily available. However, radioimmune and radiobinding assays are greatly facilitating analyses of this type. Naturally occurring 2-5A can be detected in crude trichloroacetic acid-soluble extracts, which are simple to prepare from intact cells.

Original languageEnglish
Title of host publicationInterferons - Part B
EditorsSydney Petska
PublisherAcademic Press
Pages199-208
Number of pages10
Volume79
EditionC
ISBN (Print)9780121819798
DOIs
Publication statusPublished - 1 Jan 1981
Externally publishedYes

Publication series

NameMethods in Enzymology
PublisherElsevier
ISSN (Print)0076-6879

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