MCAT Basics (from MedSchoolCoach) Separation and Purification Techniques
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Nov 26, 2025 Dive into the fascinating world of separation and purification techniques essential for the MCAT. Discover how small molecules can be separated through extraction, crystallization, and chromatography. Uncover the intricacies of protein purification, featuring SDS-PAGE and isoelectric focusing. Finally, explore methods for cell separation, including centrifugation and flow cytometry. This informative discussion sheds light on key techniques that are crucial for mastering exam content and understanding biochemical processes.
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SDS-PAGE Standardizes Charge And Shape
- SDS denatures proteins and standardizes negative charge, letting PAGE separate by molecular weight.
- Smaller proteins migrate further through the polyacrylamide gel.
Pick The Right PAGE Variant
- Use reducing SDS-PAGE to reveal subunits by breaking disulfide bonds, and non-reducing SDS-PAGE to keep subunits intact.
- Run native PAGE to preserve shape, charge, and aggregation state when needed.
Mechanisms Behind Protein Precipitation
- Protein precipitation reduces solubility to concentrate proteins via salts, pH shifts to pI, or miscible solvents.
- Removing surface charge or displacing water shells drives aggregation and precipitation.