HiTrap IMAC FF is prepacked with IMAC Sepharose 6 Fast Flow. The resin is charged with the metalion of your choice for Immobilized Metal ion Affinity Chromatography (IMAC) and subsequent purification of polyhistidine tagged proteins.
- For optimizing purification of histidine-tagged proteins when Ni²⁺ is not the best choice of metal ion
- Conveniently charge with your metal of choice
- Optimal bead size for scaling up
- High binding capacity
IMAC Sepharose FF is supplied free of metal ions. It is charged by the user with the transition metal ion of choice (e.g. Cu²⁺, Zn²⁺, Ni²⁺, or Co²⁺); these metal ions will bind to the covalently immobilized chelating ligand on the Sepharose.
IMAC Sepharose 6 Fast Flow consists of 90 µm beads of highly cross-linked agarose to which a chelating group has been covalently coupled. This chelating group can be charged with suitable metal ions, specific for your target protein of interest. The bead size of IMAC Sepharose 6 Fast Flow enables high flow properties, making the resin an excellent choice for scaling up purification at laboratory scale and purification of histidine-tagged proteins at process scale. Low metal ion leakage means that the activity of the purified protein is retained and the risk of precipitation reduced, which results in increased purity, activity, and yield of the target protein. Leakage of metal ions into the eluted target protein pool from IMAC Sepharose 6 Fast Flow is generally low under normal conditions.
IMAC Sepharose 6 Fast Flow is highly stable and compatible with a wide range of common additives. This helps maintain biological activity and increase product yield, while at the same time greatly expanding the range of suitable operating conditions.
The resin is available prepacked in HiTrap HiScreen and HiPrep column formats for convenient purification of histidine-tagged proteins at laboratory scale. Initial purification using a 1 ml HiTrap IMAC FF column can be scaled up further with the 5 ml column. Purification can be scaled up even more using 20 ml HiPrep IMAC FF 16/10 columns. Prepacked HiScreen IMAC FF with a 10 cm bed height is the format of choice for method optimization and process development.
