Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1986 Dec;83(23):8939–8942. doi: 10.1073/pnas.83.23.8939

Post-separation detection of nucleic acids and proteins by neutron activation.

R M Snapka, K Kwok, J A Bernard, O K Harling, A Varshavsky
PMCID: PMC387049  PMID: 3466168

Abstract

We describe approaches to neutron activation analysis and their application to post-separation autoradiographic detection of biological compounds. Specifically, we have extended the use of a "direct-labeling" method to the post-separation detection of DNA after gel electrophoresis and to the detection of nucleotides separated by TLC. In addition, we describe a more generally applicable "indirect-labeling" method in which separated compounds of interest are selectively bound to ligands containing highly neutron-activatable elements, such as manganese (55Mn), europium (151Eu), or dysprosium (164Dy), and then irradiated with thermal neutrons. This method is illustrated with nucleotides separated by TLC and with proteins separated by polyacrylamide gel electrophoresis. In contrast to the direct-labeling approach, the indirect-labeling method can be adapted to detect any class of substances for which a highly neutron-activatable, selectively binding ligand is available. The theoretically achievable sensitivity of the indirect-labeling method is in the attomole (10(-18) mol) range.

Full text

PDF
8939

Images in this article

8940Image
on p.8940
8940Image
on p.8940
8941Image
on p.8941
8941Image
on p.8941
8941Image
on p.8941

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BLOMSTRAND R., NAKAYAMA F. Neutron activation paper chromatographic analysis of human cerebrospinal fluid. J Neurochem. 1961 Dec;8:230–233. doi: 10.1111/j.1471-4159.1961.tb13547.x. [DOI] [PubMed] [Google Scholar]
  2. Bochner B. R., Ames B. N. Complete analysis of cellular nucleotides by two-dimensional thin layer chromatography. J Biol Chem. 1982 Aug 25;257(16):9759–9769. [PubMed] [Google Scholar]
  3. Cleaver J. E., Thomas G. H., Burki H. J. Biological damage from intranuclear tritium: DNA strand breaks and their repair. Science. 1972 Sep 15;177(4053):996–998. doi: 10.1126/science.177.4053.996. [DOI] [PubMed] [Google Scholar]
  4. Cooper P. C., Burgess A. W. Biosynthetic labeling with 32P: radiation damage to mammalian cells. Anal Biochem. 1985 Feb 1;144(2):329–335. doi: 10.1016/0003-2697(85)90125-3. [DOI] [PubMed] [Google Scholar]
  5. JOHNSON P., WEBER E. J., CARTER H. E., KROBER M. S. DETECTION OF PHOSPHOLIPIDS ON PAPER CHROMATOGRAMS BY NEUTRON ACTIVATION. J Lipid Res. 1965 Jul;6:425–426. [PubMed] [Google Scholar]
  6. Minor R. R. Cytotoxic effects of low levels of 3H-, 14C-, and 35S-labeled amino acids. J Biol Chem. 1982 Sep 10;257(17):10400–10413. [PubMed] [Google Scholar]
  7. Pollack A., Bagwell C. B., Irvin G. L., 3rd Radiation from tritiated thymidine perturbs the cell cycle progression of stimulated lymphocytes. Science. 1979 Mar 9;203(4384):1025–1027. doi: 10.1126/science.424727. [DOI] [PubMed] [Google Scholar]
  8. ROBINSON J. R. A technique for neutron activation of phosphorus compounds on paper chromatograms. Can J Biochem Physiol. 1962 Oct;40:1460–1463. [PubMed] [Google Scholar]
  9. STEIM J. M., BENSON A. A. DERIVATIVE ACTIVATION CHROMATOGRAPHY. Anal Biochem. 1964 Sep;9:21–34. doi: 10.1016/0003-2697(64)90080-6. [DOI] [PubMed] [Google Scholar]
  10. STRICKLAND E. H., BENSON A. A. Neutron activation paper chromatographic analysis of phosphatides in mammalian cell fractions. Arch Biochem Biophys. 1960 Jun;88:344–348. doi: 10.1016/0003-9861(60)90247-2. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES