FAQ-496
Can Ni-NTA resins be used to purify protein with an internal His-tag?
Yes, Ni-NTA Agarose and Superflow will bind a 6xHis-tag whether it is located internally or at the C- or N-teminal end of the protein. Note that the His-tag must be exposed for binding at the surface of the protein to allow for efficient purification under native conditions.
Related products
Ni-NTA Fast Start Kit
The Ni-NTA Fast Start Kit provides everything needed for fast, efficient purification of His-tagged proteins from cleared E.coli lysates, including prefilled Ni-NTA columns. Buffers supplied in the kit enable proteins to be purified either under native or denaturing conditions. The kit also contains an Anti-His antibody for detection of expressed His-tagged proteins.
Ni-NTA Superflow 96 BioRobot Kit
The Ni-NTA Superflow 96 BioRobot Kit provides automated purification of up to 300 µg recombinant protein from 96 samples in parallel (standard protocol, 5 ml culture volume per sample). Cleared bacterial cell lysates flow onto a filter plate preloaded with Ni-NTA Superflow by the BioRobot workstation. This unique 96-well metal-chelate affinity-chromatography module strongly and selectively binds His-tagged proteins. Ready-to-run protocols are provided for purification under native or denaturing conditions.
Ni-NTA Superflow
Ni-NTA Superflow is an affinity chromatography matrix for purifying recombinant proteins carrying a His tag.
Histidine residues in the His tag bind to the vacant positions in the coordination sphere of the immobilized nickel ions with high specificity and affinity. Cleared cell lysates are loaded onto the matrices. His-tagged proteins are bound, and other proteins pass through the matrix. After washing, His-tagged proteins are eluted in buffer under native or denaturing conditions.
Ni-NTA Spin System
Ni-NTA silica combines Ni-NTA with a macroporous silica support material optimized to suppress nonspecific hydrophobic interactions. Ni-NTA Spin Columns (His-protein purification spin columns) in the Ni-NTA Spin Kit and available separately provide Ni-NTA silica in a convenient microspin format for easy preparation of multiple samples in parallel. They provide a simple method for functional screening of engineered proteins, selection of clones expressing full-length translation products and comparison of expression levels. Each spin column can purify up to 300 µg of His-tagged protein. Like all Ni-NTA matrices, Ni-NTA spin columns can be used for one-step protein purification under native or denaturing conditions. The Ni-NTA Spin Kit is a complete kit for spin purification of His-tagged proteins. It can be automated on the QIAcube Connect (see image "QIAcube Connect").
Ni-NTA Agarose
Ni-NTA Agarose is an affinity chromatography matrix for purifying recombinant proteins carrying a His tag. Histidine residues in the His tag bind to the vacant positions in the coordination sphere of the immobilized nickel ions with high specificity and affinity. Cleared cell lysates are loaded onto the matrices. His-tagged proteins are bound, and other proteins pass through the matrix. After washing, His-tagged proteins are eluted in buffer under native or denaturing conditions