His60 Ni Superflow Cartridges using FPLC

Up to 60 mg/ml of his-tagged protein & great purity
For purification under native or denaturing conditions with low nickel ion leakage
Fast and reproducible processing on any LC system, including AKTA

His60 Ni Superflow Cartridges - Overview

His 60 Ni Superflow, the highest capacity resin to date that provides great purity, is now available in prefilled 1 ml and 5 ml cartridges for his-tagged protein purification on any LC system, such as AKTA or FPLC, or for manual purification using a syringe. The resin enables fast, easy, and reproducible chromatographic separations and can be regenerated for multiple uses. For additional information, please refer to the User Manual for His60 Nickel His-Tagged Protein Purification Cartridges.

For additional information about the resin, please see the His60 Ni Superflow Product Page

Protein Purification Principle

His60 Ni Protein Purification Resin, which is based on a modified Ni-IDA resin, provides remarkable selectivity and reliability when is it used to purify proteins containing an affinity tag of six or more consecutive histidine residues¡ªsuch as the 6xHis tag or other his tags. This technology allows one-step purification of almost any his-tagged protein from any expression system under native or denaturing conditions. The His60 Ni Superflow purification resin has low nickel ion leakage and the highest binding capacity to date (outperforming Ni-NTA and other resins).

His-Tagged Protein Purification Procedure

The purification of recombinant his-tagged proteins consists of 4 steps: cell lysis/extraction, binding, washing, and elution. His60 Nickel allows one-step protein purification under either native or denaturing conditions. The resin is compatible with multiple denaturants and detergents (Table I)¡ªor refer to the User Manual for a complete reagent compatibility table for His60 Ni resin. Purified recombinant his-tagged proteins can be eluted by the addition of 300 mM imidazole or by a reduction in pH.

Applications for Eluted His-Tag Proteins

Purified recombinant his-tagged proteins can be used for:

• Protein labeling
• Immunization to produce antibodies
• Animal studies
• Other applications

FPLC Application Data for His60 Nickel Cartridges

FPLC protein purification

Step gradient elution using His60 Ni Superflow.

6xHN-tagged AcGFP1 was purified using a 1 ml His60 Ni Superflow-packed cartridge connected to an FPLC system at a flow rate of 1 ml/min following the standard protocol described in the Recombinant His-Tagged Protein Purification User Manual. 1 ml fractions were collected and analyzed (Figure 1).

Performance analysis - comparison to Ni-NTA resin

His60 Ni Superflow resin performance was compared to the performance of Ni- NTA Superflow resin. 6xHN-AcGFP1 was purified from equivalent amounts of the same sample, following each manufacturer¡¯s protocol. Significantly higher yields and better purity of recombinant his-tagged protein were obtained using His60 Ni Superflow resin when compared to Ni-NTA resin (Figure 2).

Reproducibility of Results

His60 Ni Superflow Cartridges were tested for cartridge-to-cartridge performance. Figure 3 presents the chromatography data from three runs on three His60 Ni Superflow Cartridges, using the His60 Ni Buffer Set for protein extraction and purification. A high level of reproducibility was observed between runs, with differences of less than 5% in both protein content and fluorescence signal (data not shown).



Figure 1. Step gradient elution* results of recombinant his-tagged protein purified using His60 Ni Superflow Resin.