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

Snapka, Robert M. and Kwok, Kwan and Bernard, John A. and Harling, Otto K. and Varshavsky, Alexander (1986) Post-separation detection of nucleic acids and proteins by neutron activation. Proceedings of the National Academy of Sciences of the United States of America, 83 (23). pp. 8939-8942. ISSN 0027-8424. http://resolver.caltech.edu/CaltechAUTHORS:SNApnas86

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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.


Item Type:Article
Additional Information:Copyright © 1986 by the National Academy of Sciences. Communicated by Gerald N. Wogan, August 21, 1986. We thank Mark Solomon and Daniel Finley for comments on the manuscript and Barbara Doran for secretarial assistance. This work was supported by grants to A.V. from the National Institutes of Health (CA30367 and GM33401) and by the Nuclear Reactor Laboratory at the Massachusetts Institute of Technology. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Subject Keywords:thermal neutrons; indirect labeling; autoradiography; nucleotides; chromatography
Record Number:CaltechAUTHORS:SNApnas86
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:SNApnas86
Alternative URL:http://www.pnas.org/cgi/content/abstract/83/23/8939
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:1574
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:31 Jan 2006
Last Modified:26 Dec 2012 08:45

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