FAQ-628
Why do you recommend using Triton X for the purification of 6xHis-tagged protein?
Nonionic detergents such as Triton X-100 (0.1 - 1%) and Tween 20 (up to 2%) can be used to reduce non-specific binding of contaminating proteins due to non-specific hydrophobic or ionic interactions. They will have no effect on the binding of 6xHis-tagged protein to the Ni-NTA resin when used within the recommended concentration range.
Optimal concentrations for these additives to binding and wash buffers should be determined empirically for each purification protocol and protein.
Related products
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