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Protein determination using methylene blue in a synchronous fluorescence technique
Xiaoyu Liu1, Xia Wu, Jinghe Yang
1Key Laboratory of Colloid and Interface Chemistry (Shandong University), Ministry of Education, School of Chemistry and Chemical Engineering, Shandong University, Jinan, Shandong Province, PR China.
A novel method enhances protein detection using near-infrared fluorescence and methylene blue dedimerization. This technique offers sensitive quantification of proteins like bovine serum albumin and egg albumin.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Spectroscopy
Background:
- Methylene blue (MB) exhibits dedimerization phenomena.
- Synchronous fluorescence enhancement is a sensitive detection technique.
- Near-infrared (NIR) fluorescence offers unique spectral properties.
Purpose of the Study:
- To develop a new method for protein detection.
- To utilize the combination of NIR fluorescence and MB dedimerization.
- To investigate the fluorescence enhancement mechanism.
Main Methods:
- Developing a synchronous fluorescence enhancement method.
- Utilizing methylene blue (MB) and sodium dodecyl benzene sulfonate (SDBS).
- Analyzing bovine serum albumin (BSA) and egg albumin (EA) concentrations.
Main Results:
- Established linear relationships between fluorescence enhancement and protein concentration for BSA and EA.
- Achieved detection limits of 8.9 ng/mL for BSA and 10.0 ng/mL for EA.
- Elucidated the fluorescence enhancement mechanism involving hydrophobic interactions and complex formation.
Conclusions:
- The developed method provides sensitive and linear detection of proteins.
- The fluorescence enhancement is attributed to the formation of an MB-SDBS-protein complex and MB deaggregation.
- This approach offers a promising tool for protein quantification in analytical chemistry.
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