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Published on: June 18, 2013
A source of error in the fluorometric determination of protein in human placental microsomes
Experientia
|May 15, 1979
Abstract:
When protein is assayed in human placental microsomes by the fluorescamine procedure, 25--30% of the observed fluorescence results from labeling of phospholipids.
Insights
The fluorescamine assay for protein in human placental microsomes is affected by phospholipids. Approximately 25-30% of the fluorescence signal originates from phospholipid labeling, not just protein.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Cell Biology
Background:
- Accurate protein quantification is crucial in biological research.
- The fluorescamine assay is a common method for protein measurement.
- Human placental microsomes are complex biological samples.
Purpose of the Study:
- To investigate the specificity of the fluorescamine assay in human placental microsomes.
- To identify potential sources of interference in protein assays.
- To determine the contribution of phospholipids to fluorescence signals.
Main Methods:
- Protein assay using the fluorescamine procedure.
- Analysis of human placental microsomes.
- Fluorescence measurement.
Main Results:
- The fluorescamine assay in human placental microsomes yielded significant fluorescence from non-protein components.
- Phospholipids accounted for 25-30% of the total observed fluorescence.
- This indicates a notable interference in protein quantification.
Conclusions:
- The fluorescamine assay is not entirely specific for protein in human placental microsomes.
- Phospholipid labeling significantly impacts protein assay results.
- Consideration of lipid interference is necessary for accurate protein quantification in this sample type.
