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Protein determination in materials containing melanin.
E Vedralová1, J Borovanský, J Duchon
1Department of Biochemistry, Faculty of Medicine, Charles University, Prague, Czechoslovakia.
Journal of Biochemical and Biophysical Methods
|September 1, 1987
Summary
Determining protein content in melanin samples using the Lowry assay requires specific conditions. Melanin interference necessitates data correction, making the process complex and time-consuming.
Area of Science:
- Biochemistry
- Analytical Chemistry
Background:
- Melanin-containing samples present challenges for accurate protein quantification.
- Standard protein determination methods may be affected by melanin's optical properties.
Purpose of the Study:
- To establish prerequisites for using the Lowry procedure in melanin-containing samples.
- To develop a strategy for accurate protein determination in melanosomes.
Main Methods:
- Optimizing protein solubilization from melanosomes using sodium dodecyl sulfate (SDS) at high temperatures.
- Implementing a correction method for Folin reaction data to account for melanin interference.
- Quantifying solubilized melanin and subtracting its contribution to absorbance values.
Main Results:
- Complete protein solubilization from melanosomes required 5 hours of treatment with 3% SDS at 100°C.
- Melanin interference in the Folin reaction was significant, necessitating data correction.
- The correction for melanin interference accounted for approximately 40% of the absorbance value.
Conclusions:
- Protein determination in melanin-containing samples using the Lowry assay is feasible with data correction.
- The established correction procedure is complex and labor-intensive.
- A simpler, more efficient method for protein quantification in these samples has not yet been developed.