High Affinity Ni-Charged Resin FF (星空体育网页版)

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 星空体彩app官方. 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 FFMatrix spherical6% highly cross-linked agaroseAverage particle size90 µm (45-165 µm)Dynamic Binding capacity1≥50 mg of histidine-tagged protein /mL settled resinStorage solution 20% ethanolStorage temperaturestored at 2-8 °C; DO NOT FREEZE1 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: 3minFlow rate: 0. 333 mL/minBinding buffer: 20 mM sodium phosphate, 500 mM NaCl, 10 mM imidazole, pH 7. 4Elution 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 FFReducing agentsDenaturantsDetergentsSaltsOthers1 mM DTT6 M Gua·HCl†2% Triton X-1004 M MgCl250% glycerol20 mM β-ME       8 M Urea†  2% Tween 205 mM CaCl220% ethanol5mM TCEP-HCl——50% glycerin2 M NaCl1 mM EDTA‡10 mM reduced glutathione————100mM Na2SO460 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.