Efficient and reproducible sample preparation methods are key to successful 2-D electrophoresis. Sample preparation methods range from extraction with simple solubilization solutions to the use of complex mixtures of chaotropic agents, detergents, and reducing agents. Sample preparation can include enrichment strategies or separating protein mixtures into reproducible fractions to reduce their complexity.
Related Topics: Protein Extraction and Cleanup, Protein Fractionation and Depletion, 2-D Electrophoresis, and Troubleshooting 2-D Gels with 2-D Electrophoresis Doctor™.
An effective sample preparation procedure will:
Most protein extraction projects will require empirical determination of optimal conditions for your 2-D PAGE application. Variations in the concentrations of chaotropic agents, detergents, ampholytes, and reducing agents can dramatically affect the 2-D pattern. Find sample preparation and protein quantitation methods in the Protocols section below.
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