Key Features |
Characteristics of High Affinity Ni-Charged
Resin FF
Matrix
spherical
| 6%
highly cross-linked agarose
| Average
particle size
| 90 µm (45-165 µm)
| Dynamic
Binding capacity1
| ≥50
mg of histidine-tagged protein /mL settled resin
| Storage
solution
| 20%
ethanol
|
1 Dynamic binding capacity conditions: Sample: 5 mg/mL (histidine)6-tagged pure
proteins (Mr 43 000) in binding buffer (capacity at 10% Breakthrough) Column volume: 1
mL Retention
time: 3min Flow
rate: 0.333
mL/min Binding
buffer: 20 mM
sodium phosphate, 500 mM NaCl, 10 mM imidazole, pH 7.4 Elution
buffer: 20 mM
sodium phosphate, 500 mM NaCl, 250 mM imidazole, pH 7.4 Note: Dynamic binding capacity is
protein-dependent. Reagents Compatible with High Affinity
Ni-Charged Resin FF
Reducing agents | Denaturants
| Detergents | Salts | Others |
---|
1 mM DTT | 6 M Gua·HCl†
| 2% Triton X-100 | 4 M MgCl2 | 50% glycerol | 20 mM β-ME
| 8 M Urea†
| 2% Tween 20 | 5 mM CaCl2 | 20% ethanol | 5mM TCEP-HCl | —— | 50% glycerin | 2 M NaCl | 1 mM EDTA‡ | 10 mM reduced glutathione | —— | —— | 100mM Na2SO4 | 60 mM citrate† |
† Tested for 1
week at 40°C. ‡ The
strong chelator EDTA has been used successfully in some cases at 1 mM.
Generally, chelating agents should be used with caution (and only in the
sample, not in buffers). Any metal ion stripping may be counteracted by
addition of a small excess of MgCl2 before centrifugation/filtration of the
sample. Note that stripping effects may vary with applied sample volume. |