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Flow injection fluorometry of protein using hypochlorite-thiamine reagent
T Yokoyama1, N Nakamura, T Kinoshita
1Central Research Laboratory, SS Pharmaceutical Co. Ltd., Chiba, Japan.
Analytical Biochemistry
|January 1, 1990
Summary
This study introduces a new flow injection assay for protein quantification. The method uses thiamine to detect N-chlorides formed from peptide bonds, offering a sensitive and reproducible protein measurement.
Area of Science:
- Analytical Chemistry
- Biochemistry
Background:
- Accurate protein quantification is crucial in various scientific disciplines.
- Existing protein assays may have limitations in sensitivity or reproducibility.
Purpose of the Study:
- To develop a novel, sensitive, and reproducible flow injection assay for protein quantification.
- To utilize the reaction between N-chlorides and thiamine for protein detection.
Main Methods:
- A flow injection analysis system was employed.
- Protein samples were chlorinated using sodium hypochlorite to form N-chlorides on peptide bonds.
- Thiamine and sodium nitrite were used as fluorescence reagents to detect N-chlorides, forming fluorescent thiochrome.
- Bovine serum albumin (BSA) was used as a standard protein.
Main Results:
- The assay demonstrated high sensitivity, with a linear range from 20 ng to 2 micrograms of BSA.
- The assay showed good reproducibility.
- Fluorescence intensity was directly proportional to the protein concentration due to the number of peptide bonds.
Conclusions:
- The developed flow injection protein assay is sensitive, reproducible, and effective for quantifying protein concentrations.
- The method offers a reliable alternative for protein analysis in various applications.