ProtiFi S-Trap Mini Spin Columns for Proteomics Sample Preparation

ProtiFi S-Trap™ Mini Spin Columns are centrifugal sample-preparation columns designed for rapid protein processing prior to LC-MS/MS proteomics. The Mini format is intended for samples containing 100–300 µg of total protein and uses patented S-Trap™ technology to combine robust SDS-based protein solubilization, trapping of denatured proteins, removal of mass-spectrometry-incompatible contaminants, and enzymatic digestion directly within the column.

S-Trap™ Technology

S-Trap™ technology allows proteins to first be efficiently solubilized in an SDS-containing environment. Following reduction and alkylation, the sample is acidified and combined with a methanolic Binding/Wash Solution. Under these conditions, denatured proteins are trapped in the porous S-Trap™ matrix while contaminants are removed during washing.

Protease is then added directly to the column. Digestion takes place within the matrix, after which the resulting peptides are eluted for subsequent LC-MS/MS analysis.

Standard Workflow

1. Protein solubilization and denaturation
2. Reduction
3. Alkylation
4. Sample acidification to pH ≤1
5. Addition of methanolic Binding/Wash Solution
6. Protein trapping in the S-Trap™ Mini Spin Column
7. Washing and contaminant removal
8. Protease addition
9. In-column digestion
10. Peptide elution
11. Eluate concentration and preparation for LC-MS/MS

Technical Specifications

• Format: Centrifugal spin column
• Application: Proteomics sample preparation
• Recommended Total Protein Amount: 100–300 µg
• Reservoir Capacity: approximately 600 µL
• Standard Protein-Trapping Centrifugation: 10,000 × g for 30 s
• Maximum Centrifugation if required: up to 15,000 × g
• Washing: 400 µL Binding/Wash Solution, 10,000 × g for 30 s, repeated 3 times
• Digestion Solution Volume: 125 µL
• Recommended Protease-to-Protein Ratio: 1:10 to 1:100 (w/w)
• Current Protocol Digestion: 1–2 h at 47 °C
• First Standard Elution: 80 µL Digestion Solution
• Second Elution: 80 µL Elution Solution
• Optional Third Elution for Hydrophobic Peptides: 80 µL Elution Solution 2
• Flow Methods: centrifugation; optional vacuum or positive pressure
• Downstream Analysis: LC-MS / LC-MS/MS
• Intended Use: Research Use Only – not for use in diagnostic procedures

Available Pack Sizes

• PN C002-MINIX-0010PK – 10 S-Trap™ Mini Spin Columns
• PN C002-MINIX-0040PK – 40 S-Trap™ Mini Spin Columns
• PN C002-MINIX-0080PK – 80 S-Trap™ Mini Spin Columns