|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 (see Figure 1) . 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.|
|Key Features||Characteristics of High Affinity Ni-Charged Resin FF
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
† 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.
|Storage||Store in 20% ethanol at 2°C to 8°C. Do not freeze.|