Preparation SDS-PAGE Running Buffer
This guide describes the preparation of SDS-PAGE Running Buffer at a defined molarity for laboratory use.
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SDS-PAGE running buffer is a standard electrophoresis buffer used in Laemmli systems for protein separation under denaturing conditions.
SDS-PAGE Running Buffer – 1X Working Solution Composition
| Name | Formula | Concentration (1X) | CAS |
|---|---|---|---|
| Tris base | C₄H₁₁NO₃ | 25 mM | 77-86-1 |
| Glycine | NH₂CH₂COOH | 192 mM | 56-40-6 |
| SDS | C₁₂H₂₅O₄NaS | 0.1% (w/v) | 151-21-3 |
Applications of SDS-PAGE Running Buffer
SDS-PAGE running buffer is used for protein electrophoresis under denaturing conditions. In combination with SDS-containing sample buffer and polyacrylamide gels, it enables proteins to migrate primarily according to molecular weight by maintaining a uniform negative charge-to-mass ratio.
Preparation Tips, Sterilization, and Storage
- The pH of the solution should be 8.3 and does not require any adjustment.
- SDS readily generates foam. When adding water, allow it to flow slowly down the wall of the container to minimize foaming, and avoid vigorous stirring.
- SDS may form flocculent precipitates at low temperatures. Ensure it is fully dissolved before use.
- Sterilization: usually not required; choose as needed.
- Storage: room temperature.
