Ni-NTA Agarose 100
PureCube 100 Ni-NTA Agarose
The His tag is the most widely used affinity tag due to its small size, low immunogenicity, and versatility under native or denaturing conditions, as well as in presence of detergents and many other additives. Cube Biotech introduces the novel PureCube 100 Ni-NTA Agarose, based on high quality crosslinked agarose with 100 µm mean diameter, providing high flow rates in batch and FPLC applications. Due to the excellent flow properties of PureCube 100 Agarose, combined with the high capacity NTA ligand, high dynamic yield and protein purity can be achieved. Even at flow rates of 7 ml/min, more than 20 mg/ml his-tagged protein could be purified. In batch setups, protein yields were at least 80 mg/ml (Fig.1) PureCube 100 Ni-NTA Agarose shows high affinity to his-tagged proteins. Recovery rates of 85-nearly 100% were achieved, even with low concentrated protein samples (Fig.2). PureCube 100 Ni-NTA Agarose is provided as a 50% suspension. To detect His-tagged proteins in Western Blot experiments, Cube Biotech offers the highly specific PentaHis antibody.
Why PureCube 100 Ni-NTA Agarose?
- 100 µm agarose beads for high flow rates in batch and FPLC
- High binding capacity (up to 80 mg/mL) and high purity even at high flow rates
- Excellent recovery even with low concentrated samples
- Stable in 10 mM DTT and 1 mM EDTA. Need to use higher concentrations?
The features of Ni-NTA 100 Resins in detail
Fig. 1: High dynamic binding capacity of PureCube 100 Ni-NTA Agarose. GFP was spiked into E.coli lysates and purified on a 1 mL PureCube Cartridge filled with PureCube 100 Ni-NTA Agarose at flow rates from 1 to 7 ml/min, and in a gravity flow batch procedure. Left: Flow diagram, Right: SDS-PAGE analysis of elution fractions 1-4 (E1-4).
Fig.2: Excellent protein recovery with PureCube 100 Ni-NTA Agarose. GFP was spiked into E.coli lysates in defined quantities (10, 5, 2.5, 1.25, and 0.625 mg) and purified on a 1 mL PureCube Cartridge filled with PureCube 100 Ni-NTA Agarose. Recovered GFP was quantified by absorption at 488 nm.