研究目的
To develop a label-free, ultra-sensitive fluorescence method for detecting cytochrome c (Cyt c) and applying it to natural drug screening targeted for Cyt c.
研究成果
The DNA-AgNCs@tween 80 nanoprobe is highly sensitive, selective, and biocompatible for Cyt c detection, enabling effective monitoring of Cyt c release during cell apoptosis and application in natural drug screening, with compound J5 identified as a potential anti-colon cancer drug.
研究不足
The concrete mechanism of fluorescence enhancement by tween 80 should be further explored. The method's applicability may be limited to specific cell types and conditions, and further validation in more complex biological systems is needed.
1:Experimental Design and Method Selection:
The study designed a fluorescence-based detection system using DNA-templated silver nanoclusters (DNA-AgNCs) enhanced with tween 80 (DNA-AgNCs@tween 80) as a nanoprobe for Cyt c detection, based on fluorescence quenching via electron transfer.
2:Sample Selection and Data Sources:
Cyt c from horse heart, HCT-116 and BGC-823 tumor cell lines, natural compounds J5 and S6 extracted from plants, and other biomolecules for selectivity tests.
3:List of Experimental Equipment and Materials:
DNA oligonucleotide, sodium borohydride, silver nitrate, phosphate buffers, tween 80, Cyt c, trypsin, ATP, BSA, amino acids, cell culture media, MTT solution, DMSO, and various instruments including F-2500 Spectrophotometer, UV-1800 Ultraviolet spectrophotometer, Zeta potentiometer, FACScan cytometer.
4:Experimental Procedures and Operational Workflow:
Synthesis of DNA-AgNCs@tween 80, optimization of temperature, pH, and incubation time, Cyt c assay with fluorescence measurement, cell culture and MTT assay for cytotoxicity, detection of released Cyt c in cell medium, and flow cytometry for apoptosis analysis.
5:Data Analysis Methods:
Fluorescence intensity measurements, linear regression for concentration relationships, statistical analysis of cell viability and apoptosis rates.
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