Ni-NTA Superflow (25 ml)

From QIAGEN, Ni-NTA Superflow is an affinity chromatography matrix designed for purifying recombinant proteins carrying a His tag. Histidine residues in the His tag bind to vacant coordination positions on immobilized nickel ions with high specificity and affinity. Cleared cell lysates are loaded onto the matrix; His-tagged proteins bind while other proteins pass through. After washing, bound proteins are eluted in buffer under native or denaturing conditions.

The matrix combines Ni-NTA coupled to Superflow resin, delivering superior mechanical stability, outstanding flow characteristics, and high dynamic binding capacity. It enables one-step purification of 6xHis-tagged proteins using high flow rates and pressures, making it suitable for production-scale and FPLC applications. The system can be used with any expression system, including baculovirus, mammalian cells, yeast, and bacteria. Purification follows four stages: cell lysis, binding, washing, and elution, and does not depend on the 3-dimensional structure of the protein or 6xHis tag, allowing purification from dilute solutions and crude lysates.

Strong denaturants and detergents can be used for solubilization and purification of receptors, membrane proteins, and inclusion body proteins. Purified proteins are eluted under mild conditions by adding 100–250 mM imidazole as a competitor or by reducing the pH. Specifications include a bead size of 60–160 µm, a binding capacity of up to 50 mg/mL, large-scale manual processing for 1–24 samples per run, and compatibility with both batch and column purification by gravity flow or automated operation. A Drug Master File (DMF) is available for regulatory support. The NTA ligand binds nickel through four chelation sites, preventing nickel-ion leaching and yielding higher binding capacity and purity than three-site chelation systems.