Equilibration of 1 mL cup. Amersham Pharmacia HiTrap Phenyl Sepharose Column
1. Wash column with 5 mL elution buffer (1ml min -1)
2. Wash column with 10 mL wash buffer
Add sample
1. Apply sample (previously resuspended in start buffer)
2. Wash with 5 mL wash buffer
3. Elute with 10 mL elution buffer
Regenerate column
1. Wash with 5 mL MQH2O
2. Wash with 5 mL wash buffer
Add new sample
Start Buffer
500 mM NaCl
10 mM CaCl2
20mM HEPES pH 7.5
Wash Buffer
500 mM NaCl
1 mM CaCl2
20 mM HEPES pH 7.5
Elution Buffer
500 mM NaCl
1 mM EDTA
20 mM HEPES pH 7.5