Description
GenScript High
Affinity Ni-Charged Resin FF is an 6% highly cross-linked agarose medium
covalently coupled to a chelating agent that binds Ni2+ by four
coordination sites for high-affinity purification of polyhistidine-tagged
recombinant proteins.
High Affinity Ni-Charged Resin FF has low Ni2+ leakage,
high protein-binding capacity and stability, and is compatible with a wide
range of additives used in protein purification. This makes High Affinity Ni Charged Resin FF the
excellent choice for high performance purification of polyhistidine-tagged
proteins.
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
| | Storage temperature | stored at 2-8 °C; DO NOT FREEZE |
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.
|
Figure 1. Schematic diagram of the chemical structure of Ni IDA Resin, Ni IDA Resin FF, Hi affinity Ni-Charged Resin and Hi affinity Ni-Charged Resin FF
For research and manufacturing use. Direct human use, including taking
orally and injection are forbidden.
