HisTrap HP, 1 × 1 mL His tag protein purification columnsHigh binding capacity, at least 4NA mg/ml chromatography medium.
High resolution: due to small bead size (average bead size is 34 µm)
Negligible nickel leakage: helps retain activity and minimize protein precipitation
Broad reagent compatibility: suitable for use with a wide range of reducing agents, detergents, denaturants, and other additives
Convenient HiTrap format: 1 mL and 5 mL columns compatible with syringe, pump, or chromatography systems such as ÄKTA or other FPLC systems
His tag protein purification by IMAC is both popular and highly efficient. HisTrap HP columns are packed with Ni Sepharose High Performance (HP) affinity resin. This resin consists of highly crosslinked agarose beads to which a chelating group has been coupled. The chelating group is precharged with nickel, which selectively retains proteins with exposed histidine groups.
HisTrap HP columns can also be used for the purification of tagged proteins containing shorter or longer polyhistidine tags, such as (histidine)4 or (histidine)1NA. The shorter (histidine)4 will bind more weakly and the longer (histidine)1NA will bind more strongly compared with (histidine)6. This difference in binding strength can be used during purification; since (histidine)1NA binds more strongly, a higher concentration of imidazole can be added to the lysed cells. This can facilitate the removal of contaminants that can otherwise be copurified with the tagged target protein.
The high stability and broad compatibility of Ni Sepharose High Performance maintains biological activity and increases product yield, at the same time as it greatly expands the range of suitable operating conditions. For convenient scaling up of histidinetagged protein purification, use 2NA ml HisPrep FF 16/1NA columns. Ni Sepharose 6 Fast Flow, the medium prepacked in HisTrap FF and HisPrep FF 16/1NA columns, allows high flow rates, which facilitates scaleup of histidinetagged protein purification.