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Colorimetric micromethod for protein determination with erythrosin B
1Kraft General Foods, Tarrytown, New York 10591.
Analytical Biochemistry
|July 1, 1994
Summary
A new protein assay using erythrosin B offers a stable, sensitive, and reproducible method for quantifying proteins in solution. This erythrosin assay provides rapid results, especially at elevated temperatures, making it suitable for routine analysis.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Spectroscopy
Background:
- Accurate protein quantification is crucial for various biological and chemical analyses.
- Existing protein assays have limitations in sensitivity, reproducibility, or interference.
Purpose of the Study:
- To develop and characterize a novel quantitative protein determination method using erythrosin B.
- To evaluate the performance of the erythrosin B assay for routine analytical applications.
Main Methods:
- Proteins were reacted with erythrosin B to form a colored chromophore.
- Absorbance was measured at 545 nm to quantify protein concentration.
- Assay performance was evaluated at room temperature and elevated temperatures (90-95°C).
Main Results:
- The reaction yields a stable, highly colored chromophore with maximal absorbance at 545 nm.
- The assay demonstrated high sensitivity (2-14 µg/ml protein) and good reproducibility.
- Faster reaction times (1.5-2 min) and increased sensitivity were observed at 90-95°C.
- The assay showed limited interference from common reagents and low protein-to-protein variability.
Conclusions:
- The erythrosin B assay is a sensitive, reproducible, and rapid method for protein quantification in solution.
- Its characteristics make it suitable for routine analytical purposes, potentially overcoming limitations of other assays.