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Quantitative analysis of electrophoretically separated proteins using Coomassie blue
The Japanese Journal of Experimental Medicine
|August 1, 1978
Summary
This study developed a sensitive electrophoresis method for quantifying rat albumin and gamma-globulin. The technique accurately measures protein levels, revealing circadian rhythm impacts on urinary gamma-globulin excretion in rats.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Proteomics
Background:
- Quantifying protein levels is crucial in biological research.
- Existing methods may lack sensitivity or require large sample volumes.
- Developing precise techniques for specific proteins like albumin and gamma-globulin is essential.
Purpose of the Study:
- To establish a sensitive and linear electrophoresis method for quantifying rat albumin and gamma-globulin.
- To demonstrate the method's applicability in studying physiological variations.
Main Methods:
- Capillary electrophoresis using a 4-40% continuous gradient polyacrylamide gel.
- Quantification of 125I-labelled rat albumin and gamma-globulin.
- Analysis of protein amounts, densitometric areas, and radioactivity using double logarithm plots.
Main Results:
- A linear relationship was established between protein amounts and densitometric areas for both albumin and gamma-globulin.
- Radioactivity measurements also showed a linear correlation with protein amounts.
- The method successfully detected circadian rhythm-induced changes in urinary gamma-globulin excretion in rats.
Conclusions:
- The developed capillary electrophoresis method offers a sensitive and reliable approach for quantifying rat albumin and gamma-globulin.
- This technique is valuable for detecting physiological changes, such as circadian variations in urinary protein excretion.