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Artifacts associated with 2-mercaptoethanol upon high resolution two-dimensional electrophoresis.
Analytical Biochemistry
|June 1, 1984
Summary
2-mercaptoethanol in protein sample preparation for 2D electrophoresis can cause silver staining artifacts. These artifacts, including spots, bands, and streaks, likely originate from contaminants released by 2-mercaptoethanol.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Molecular Biology
Background:
- High-resolution two-dimensional electrophoresis (2D-PAGE) is a powerful technique for protein separation.
- Silver staining is a sensitive method for detecting proteins in 2D-PAGE gels.
- 2-mercaptoethanol is commonly used as a reducing agent in protein sample preparation for 2D-PAGE.
Purpose of the Study:
- To investigate artifacts observed in 2D-PAGE silver-stained patterns.
- To identify the source and nature of these artifacts potentially related to 2-mercaptoethanol.
Main Methods:
- High-resolution two-dimensional electrophoresis (2D-PAGE).
- Silver staining for protein detection.
- Analysis of artifact characteristics (isoelectric point, electrophoretic mobility, molecular weight).
Main Results:
- Observed artifacts include discrete spots, horizontal bands (MW 54,000 and 68,000), and vertical streaks.
- Artifacts appeared in silver-stained 2D-PAGE patterns.
- Evidence suggests artifacts are not from 2-mercaptoethanol itself but from released contaminants.
Conclusions:
- 2-mercaptoethanol in sample preparation can introduce artifacts in silver-stained 2D-PAGE.
- These artifacts may arise from contaminants released from common electrophoretic system components.
- Further investigation is needed to pinpoint the exact source of these contaminants.