Tris Tricine SDS (10X): High-Resolution Separation of Small Proteins and Peptides
Standard discontinuous SDS-PAGE employing Tris-Glycine as the tank buffer is highly effective for resolving proteins down to approximately 15 kDa. However, below this threshold, smaller proteins and peptides fail to destack from the SDS micelles, often migrating with the buffer front and resulting in poor resolution or loss of data. The Geneflow Tris-Tricine SDS (10X) buffer is specifically engineered to address this limitation, providing a robust solution for researchers requiring clear, distinct bands for low molecular weight targets.
Based on the specialised system developed by Schagger and von Jagow, this 10X concentrate offers a streamlined alternative to traditional peptide electrophoresis. By substituting the standard Tris-Glycine cathode (upper) tank buffer with this Tris-Tricine-SDS formulation, the resolution range of standard SDS-PAGE is extended down to 5 kDa. Crucially, this system is designed to be compatible with the standard Laemmli gel and buffer chemistry, allowing for enhanced resolution with minimal alteration to existing laboratory protocols.
Key Benefits:
-
- Extended Resolution Range: Detect smaller targets. Enables the clear separation and resolution of proteins and peptides as small as 5 kDa.
- Seamless Integration: Minimal protocol changes. Simply substitute the Tris-Glycine buffer in the cathode tank with Tris-Tricine-SDS; no changes to the gel formulation are required.
- High Precision: Sharper peptide bands. Optimised ion mobility ensures that low molecular weight proteins destack efficiently from SDS micelles for publication-quality results.
- Time Efficiency: Ready-to-dilute 10X format. Eliminates the need for time-consuming reagent weighing and mixing, reducing the potential for batch-to-batch variability.
Technical Insights
Optimised Ion Mobility for Peptide Stacking
The effectiveness of the Tris-Tricine system lies in the electrophoretic mobility of the tricine ion compared to the glycine ion. Tricine has a lower pKa and different mobility characteristics, which creates a more effective stacking environment for small proteins. This ensures that even the smallest fragments are compacted into a tight band before entering the resolving gel, preventing the “smearing” typically associated with small proteins in glycine-based systems.
Specifications
- Concentration: 10X (Dilute 1:9 with deionised water).
- Application: SDS-PAGE Cathode (Upper) Tank Buffer.
- Resolution Limit: Proteins and peptides down to 5 kDa.
- Compatibility: Compatible with standard Laemmli gel systems.
- Storage: Store at Room Temperature (15°C to 25°C).
- Shelf Life: 12 months from date of manufacture.