cOmplete His-Tag Purification Resin
pre-charged with Ni2+, ready-to-use
In contrast to other available resins, which are primarily based on NTA or IDA chelator chemistry, this resin tolerates buffers that contain EDTA. Because EDTA is a known inhibitor of metalloproteases - which are frequently present in any cell type - this chromatography material provides the option to protect your target protein by supplementing your buffers with EDTA.This resin therefore allows the easy purification of your target protein, while using an effective protection against degradation. The His-Tag binding principle allows a standard one-step isolation procedure, generally for all low- and high-molecular weight proteins. Using a special strong-binding Ni2+ chelator, minimal contamination of your fractions with metal ions is ensured, safeguarding your downstream applications.ApplicationThe most common technique in affinity purification of proteins involves engineering a sequence of 6 to 14 histidines into the N- or C-terminal (or even on an exposed loop) of the protein. Such polyhistidine stretches bind strongly to divalent metal ions such as nickel and cobalt. This effect can be used to separate proteins. Metal ions can be immobilized on a matrix using a chelator, which still allows the ion to interfere with the polyhistidine tag of the protein. When these his-tagged proteins are passed through a column containing immobilized metal ions, the proteins bind via the tag to the column. Nearly all untagged proteins pass through the column. The protein is released from the column by elution with either imidazole, which competes with the polyhistidine tag for binding to the column, or by a decrease in pH, which decreases the affinity of the tag for the resin. While this procedure is generally used for the purification of recombinant proteins with an engineered affinity tag, it can also be used for natural proteins with an inherent affinity for divalent cations.
