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@inbook{1844939,
author = {Fulneček, Jaroslav and Říha, Karel},
address = {TOTOWA},
booktitle = {RNA REMODELING PROTEINS},
doi = {http://dx.doi.org/10.1007/978-1-0716-0935-4_7},
keywords = {Protein-induced fluorescence enhancement (PIFE); Cy3; RNA; DNA; Protein; Binding assay},
howpublished = {elektronická verze "online"},
language = {eng},
location = {TOTOWA},
isbn = {978-1-0716-0934-7},
pages = {109-117},
publisher = {HUMANA PRESS INC},
title = {High-Throughput Protein-Nucleic Acid Interaction Assay Based on Protein-Induced Fluorescence Enhancement},
url = {https://link.springer.com/protocol/10.1007/978-1-0716-0935-4_7#citeas},
year = {2021}
}
TY - CHAP
ID - 1844939
AU - Fulneček, Jaroslav - Říha, Karel
PY - 2021
TI - High-Throughput Protein-Nucleic Acid Interaction Assay Based on Protein-Induced Fluorescence Enhancement
VL - 2 EDITION
PB - HUMANA PRESS INC
CY - TOTOWA
SN - 9781071609347
KW - Protein-induced fluorescence enhancement (PIFE)
KW - Cy3
KW - RNA
KW - DNA
KW - Protein
KW - Binding assay
UR - https://link.springer.com/protocol/10.1007/978-1-0716-0935-4_7#citeas
N2 - Molecular processes involved in gene expression encompass multitudes of interactions between proteins and nucleic acids. Quantitative description of these interactions is crucial for delineating the mechanisms governing transcription, genome duplication, and translation. Here we describe a detailed protocol for the quantitative analysis of protein-nucleic acid interactions based on protein-induced fluorescence enhancement (PIFE). While PIFE has mainly been used in single-molecule studies, we modified its application for bulk measurement of protein-nucleic acid interactions in microwell plates using standard fluorescent plate readers. The microwell plate PIFE assay (mwPIFE) is simple, does not require laborious protein labeling, and is high throughput. These properties predispose mwPIFE to become a method of choice for routine applications that require multiple parallel measurements such as buffer optimization, competition experiments, or screening chemical libraries for binding modulators.
ER -
FULNEČEK, Jaroslav and Karel ŘÍHA. High-Throughput Protein-Nucleic Acid Interaction Assay Based on Protein-Induced Fluorescence Enhancement. Online. In \textit{RNA REMODELING PROTEINS}. TOTOWA: HUMANA PRESS INC, 2021, p.~109-117. 2 EDITION. ISBN~978-1-0716-0934-7. Available from: https://dx.doi.org/10.1007/978-1-0716-0935-4\_{}7.
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