ProtoGel Sample Prep Kit: Optimise Protein Samples for SDS-PAGE
The ProtoGel Sample Prep Kit provides a universal, convenient method for concentrating and optimising protein samples prior to SDS-PAGE analysis. Dilute samples or those containing interfering substances such as high salt concentrations often result in “smiling” gels, blurred bands, and poor resolution. This kit eliminates these common artefacts by selectively precipitating proteins while leaving behind contaminants that disrupt electrophoretic mobility.
Utilising a proprietary two-reagent system, the kit effectively captures proteins in a precipitant complex, allowing for the removal of the supernatant containing salts and other soluble impurities. Following a simple acetone solubilisation and ethanol wash steps, the result is a pure protein pellet ready for resuspension in loading buffer. This process ensures reproducible migration and excellent visualisation, even for samples that were previously too dilute or contaminated for analysis.
Key Benefits:
- High-Yield Concentration: Recover dilute proteins. Effectively concentrates proteins with a lower limit of reproducible recovery around 100ng in 40µl.
- Contaminant Removal: Sharper bands. Removes high salt concentrations and urea that cause gel distortion, ensuring crisp, straight bands.
- Broad Compatibility: Versatile application. Successfully precipitates intact proteins in the 10kD to 200kD molecular weight range within a pH range of 6 to 8.
- Simple Protocol: Fast workflow. Uses a straightforward precipitation, dissolution, and wash method that prepares samples for loading in under an hour.
Technical Insights
The ProtoGel Sample Prep Kit operates via a selective precipitation mechanism. When Reagents A and B are added to the sample, they form a matrix that entraps the protein. This complex is collected via centrifugation, allowing the removal of the original buffer containing interfering agents. The matrix is then dissolved using acetone, leaving a tight protein pellet that is washed with 70% ethanol to ensure purity.
Compatibility Notes: While robust against most biological salts, very high salt concentrations (e.g., saturated NaCl) may require prior dilution. Users should note that SDS or non-ionic surfactants (such as Tween or Triton™) above 1% concentration can reduce recovery. Additionally, the kit is incompatible with deoxycholate, thiocyanate salts, or perchlorate salts.
Specifications
- Product Name: ProtoGel Sample Prep Kit
- Catalogue Number: A2-0071 (Manufacturer Ref: EC-884)
- Pack Size: 100 Preparations
- Molecular Weight Range: 10kD – 200kD
- Sensitivity: Recovers down to ~100ng protein
- pH Tolerance: pH 6.0 – 8.0
- Incompatibilities: Deoxycholate, Thiocyanate/Perchlorate salts, >1% SDS/Tween/Triton