A new electrophoretic-autoradiographic method for the visual detection of phosphotransferases

Jay A. Tischfield, H. P. Bernhard, Frank H. Ruddle

Research output: Contribution to journalArticlepeer-review

45 Scopus citations

Abstract

In this paper we present the details of a slab acrylamide, lanthanum precipitation, autoradiographic technique and show it to be useful for the visualization of at least four different enzymes. We believe that with appropriate separation conditions and reaction mixtures this technique could be extended to a larger number of enzymes, theoretically all those whose isotopically labeled product could be specifically precipitated within the matrix of a polyacrylamide gel. It will be interesting to use this technique with other gel buffer systems, particularly those with a lower pH that have recently been reported (21). In addition, it might be useful to combine it with isoelectric focusing in slab gels (10). The technique would appear to be particularly useful for phosphotransferases and, to date, it has been applied to thymidine kinase and adenosine kinase with encouraging results. Work with other enzymes, and adapting the technique to starch gels, is also in progress.

Original languageEnglish (US)
Pages (from-to)545-554
Number of pages10
JournalAnalytical Biochemistry
Volume53
Issue number2
DOIs
StatePublished - Jun 1973
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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