Fluorometric determination of the CCAAT/enhancer binding protein alpha by using gold nanoparticles and a labeled protein-binding DNA.
- Resource Type
- Article
- Authors
- Ma, Jiehua; Li, Jinlong; Cui, Xianwei; You, Lianghui; Li, Yun; Wen, Juan; Ji, Chenbo; Guo, Xirong
- Source
- Microchimica Acta. Jan2020, Vol. 187 Issue 1, p1-7. 7p.
- Subject
- *GOLD nanoparticles
*FLUORESCENCE quenching
*DNA
*FLUORESCENT probes
*DETECTION limit
*GENE enhancers
- Language
- ISSN
- 0026-3672
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. [ABSTRACT FROM AUTHOR]