High-Throughput Protein-Nucleic Acid Interaction Assay Based on
Protein-Induced Fluorescence Enhancement

C 2021

High-Throughput Protein-Nucleic Acid Interaction Assay Based on Protein-Induced Fluorescence Enhancement

FULNEČEK, Jaroslav a Karel ŘÍHA

Základní údaje

Originální název

High-Throughput Protein-Nucleic Acid Interaction Assay Based on Protein-Induced Fluorescence Enhancement

Autoři

FULNEČEK, Jaroslav a Karel ŘÍHA

Vydání

TOTOWA, RNA REMODELING PROTEINS, od s. 109-117, 9 s. 2 EDITION, 2021

Nakladatel

HUMANA PRESS INC

Další údaje

Jazyk

angličtina

Typ výsledku

Kapitola resp. kapitoly v odborné knize

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Forma vydání

elektronická verze "online"

Odkazy

URL

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14740/21:00119726

Organizační jednotka

Středoevropský technologický institut

ISBN

978-1-0716-0934-7

Klíčová slova anglicky

Protein-induced fluorescence enhancement (PIFE); Cy3; RNA; DNA; Protein; Binding assay

Štítky

rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 5. 5. 2022 15:22, Mgr. Pavla Foltynová, Ph.D.

Anotace

V originále

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.

Návaznosti

EF15_003/0000479, projekt VaV

Název: Regulace rostlinné meiózy

GA19-21961S, projekt VaV

Název: Chemická modulace aktivity Ku komplexu a využití v editaci genomů.

Investor: Grantová agentura ČR, Chemická modulace aktivity Ku komplexu a využití v editaci genomů

Citovat

FULNEČEK, Jaroslav a Karel ŘÍHA. High-Throughput Protein-Nucleic Acid Interaction Assay Based on Protein-Induced Fluorescence Enhancement. Online. In RNA REMODELING PROTEINS. TOTOWA: HUMANA PRESS INC, 2021, s. 109-117. 2 EDITION. ISBN 978-1-0716-0934-7. Dostupné z: https://doi.org/10.1007/978-1-0716-0935-4_7.

@inbook{1844939,
   author = {Fulneček, Jaroslav and Říha, Karel},
   address = {TOTOWA},
   booktitle = {RNA REMODELING PROTEINS},
   doi = {https://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 a 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, s.~109-117. 2 EDITION. ISBN~978-1-0716-0934-7. Dostupné z: https://doi.org/10.1007/978-1-0716-0935-4\_{}7.

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