Bio-Rad Nuvia™ IMAC Resin - Manual (Page 33)

Nuvia IMAC Ni-Charged Resin 29
1. Strip metal ions.
Wash with 10 column volumes of 50 mM sodium phosphate,
0.3 M NaCl, and 0.05–0.5 M EDTA, pH 7.5.
2. Wash the column with one of the following solutions at
2 ml/min:
a. 1% acetic acid
• This solution may be used as a cleaning, sanitizing,
and storage solution with Nuvia IMAC resins
• 10–15 min exposure time
• Rinse with 10 column volumes of distilled water
b. 2 M NaCl (removes ionic contaminants)
• 10–15 min exposure time
• Rinse with 10 column volumes of distilled water
c. 1 M NaOH up to 3 hr (removes precipitated, hydrophobic,
and lipoproteins)
• Exposure time is usually 1–3 hr
• Rinse with 10 column volumes of distilled water
d. 70% ethanol or 30% isopropyl alcohol (removes
precipitated, hydrophobic, and lipoproteins)
• 15–20 min exposure time
• Alternatively, use 0–30% gradient isopropyl alcohol
over 5 column volumes, followed by 2 column
volumes 30% isopropyl alcohol
• Rinse with 10 column volumes of distilled water
3. Remove cleaning solution(s) from column by rinsing with 10
column volumes of binding buffer.
• Rinse column with 50 mM sodium phosphate,
0.3 M NaCl, pH 8
• Ensure the eluate is ~pH 8 and the UV signal has returned
to baseline
4. Recharge the column with metal ion of choice.
• Refer to Section 6, Immobilizing Metal Ions