Zhang, Zhan-Ren, Palfrey, David, Nagel, David A., Lambert, Peter A., Jessop, Robert A., Santos, Albert F. and Hine, Anna V. (2003). Fluorescent microplate-based analysis of protein-DNA interactions I:immobilized protein. Biotechniques, 35 (5), pp. 980-986.
Abstract
A simple protein-DNA interaction analysis has been developed using a high-affinity/high-specificity zinc finger protein. In essence, purified protein samples are immobilized directly onto the surface of microplate wells, and fluorescently labeled DNA is added in solution. After incubation and washing, bound DNA is detected in a standard microplate reader. The minimum sensitivity of the assay is approximately 0.2 nM DNA. Since the detection of bound DNA is noninvasive and the protein-DNA interaction is not disrupted during detection, iterative readings may be taken from the same well, after successive alterations in interaction conditions, if required. In this respect, the assay may therefore be considered real time and permits appropriate interaction conditions to be determined quantitatively. The assay format is ideally suited to investigate the interactions of purified unlabeled DNA binding proteins in a high-throughput format.
Publication DOI: | https://doi.org/10.2144/03355st06 |
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Divisions: | College of Health & Life Sciences > Aston Pharmacy School College of Health & Life Sciences > School of Biosciences College of Health & Life Sciences > Chronic and Communicable Conditions College of Health & Life Sciences College of Health & Life Sciences > School of Biosciences > Cellular and Molecular Biomedicine Aston University (General) |
Additional Information: | Creative Commons Attribution Non-Commercial No Derivatives License |
Uncontrolled Keywords: | protein-DNA interaction,zinc finger protein,DNA,Clinical Biochemistry,General Biochemistry,Genetics and Molecular Biology |
Publication ISSN: | 1940-9818 |
Last Modified: | 02 Dec 2024 08:05 |
Date Deposited: | 19 Aug 2019 08:50 |
Full Text Link: | |
Related URLs: |
http://www.scop ... tnerID=8YFLogxK
(Scopus URL) https://www.fut ... cbi.nlm.nih.gov (Publisher URL) |
PURE Output Type: | Article |
Published Date: | 2003-11 |
Authors: |
Zhang, Zhan-Ren
Palfrey, David Nagel, David A. ( 0000-0002-9055-1775) Lambert, Peter A. ( 0000-0002-8243-2741) Jessop, Robert A. Santos, Albert F. Hine, Anna V. ( 0000-0003-4065-831X) |
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