Protocol Stripping and Recharging HisTrap Column

Stripping and recharging HisTrap column

Notes: HisTrap columns need to be stripped and recharged after five to seven purifications.

Materials

0.1 M sodium phosphate buffer (pH 7.4)

3.1 g NaH2PO4-H2O

10.9 g Na2HPO4

bring up the volume to 1L

Stripping

Stripping buffer

20 mM sodium phosphate, pH 7.4

0.5 M NaCl

50 mM EDTA

Binding Buffer

20 mM sodium phosphate, pH 7.4

0.5 M NaCl

5 mM imidazole

Procedure

1. Strip the column by washing it with 5-10 CV stripping buffer.

2. Wash with 5-10 CV binding buffer.

3. Wash with 5-10 CV ddH2O.

Recharging

Recharging buffer

0.1 M NiSO4 in ddH2O

Procedure

1. Load 2.5 mL of recharging buffer

2. Wash with 5 CV ddH2O

3. 5 CV binding buffer