Reagents

• Equilibration Buffer is 50mM Tris, 5mM EDTA pH 8.5
• Binding Buffer is 1-10mg/ml peptide in Equilibration Buffer
• Blocking Buffer is 50mM Cysteine in Equilibration Buffer
• Wash Solution is 1M NaCl
• Storage Solution is 0.05% Sodium Azide - TOXIC

Sample Preparation

• Have all reagents at room temperature
• Choose peptide so that it has a free cysteine

Ligand Coupling

• Equilibrate column with 6 volumes Equilibration Buffer, drain
• Add 1ml Binding buffer
• Cap column and mix by end-over-end rocking 15 minutes
• Incubate for additional 30 minutes without mixing
• Drain then wash with 3 volumes Equilibration buffer

Blocking

• Add 1ml of Blocking Buffer per 1ml of Sulfolink Gel
• Cap column and mix by end-over-end rocking 15 minutes
• Incubate for additional 30 minutes without mixing
• Drain then wash column with 16 volumes Sash Solution
• Drain then wash column with 16 volumes Storage Solution
• Store at 4°C until ready to use