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Fluorometric determination of the CCAAT/enhancer binding protein alpha by using gold nanoparticles and a labeled protein-binding DNA

Authors
  • Ma, Jiehua1
  • Li, Jinlong2
  • Cui, Xianwei1
  • You, Lianghui1
  • Li, Yun1
  • Wen, Juan1
  • Ji, Chenbo1
  • Guo, Xirong1
  • 1 Women’s Hospital of Nanjing Medical University, Nanjing Maternity and Child Health Care Hospital, Nanjing, 210004, People’s Republic of China , Nanjing (China)
  • 2 Nanjing University of Chinese Medicine, Nanjing, 210003, People’s Republic of China , Nanjing (China)
Type
Published Article
Journal
Microchimica Acta
Publisher
Springer-Verlag
Publication Date
Dec 05, 2019
Volume
187
Issue
1
Identifiers
DOI: 10.1007/s00604-019-4025-1
Source
Springer Nature
Keywords
License
Yellow

Abstract

A method is described for the determination of the CCAAT/enhancer binding protein alpha (C/EBPα) which is a regulator in adipocyte differentiation. The method is based on quenching of the red fluorescence (with excitation/emission maxima at 548/562 nm) of Cy3-labeled DNA if it becomes adsorbed on positively charged gold nanoparticles (AuNPs). Fluorescently labeled dsDNA that can bind C/EBPα is introduced as a fluorescent probes. The dsDNA is electrostatically adsorbed on the positively charged AuNPs to quench their fluorescence. In the presence of C/EBPα, it will bind dsDNA which then diffuses away. The fluorescence of the AuNPs becomes restored. The fluorescent signal increases linearly in the 0.05 to 600 ng·mL−1 μM C/EBPα concentration range, and the detection limit is 29 pg·mL−1. The method is specific and was applied to analyze cell lysates and in-situ. Graphical abstractSchematic representation of a fluorometric method for determination of the CCAAT/enhancer binding protein alpha (C/EBPα). Fluorescently labeled dsDNA that can bind C/EBPα is introduced as a fluorescent probes. The dsDNA is electrostatically adsorbed on the positively charged AuNPs to quench their fluorescence. In the presence of C/EBPα, it will bind dsDNA which then diffuses away. The fluorescence of the AuNPs becomes restored.

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