ROCHE Complete His-tag Purification Column 1 X 5mL

Stock Code: 2795338
Manufacturer Part No: 6781535001

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Descriptions

Compatibility

Compatibility for long term storage: 20% ethanol, pH 4.0 to pH 9.0 Compatibility during chromatography: The resin is compatible with 10mM EDTA, 10mM DTT during the purification (1 hour incubation), 6M guanidinium-HCl, 8M urea, pH 2.0 to pH 14.0. Compatibility during cleaning: 4% SDS Form cOmplete His-Tag Purification Resin filled in columns, pre-charged with Ni²⁺ stored in 20% ethanol.

Features and Benefits

cOmplete His-Tag Purification Columns are available in 2 sizes and prepacked with cOmplete His-Tag Purification Resin. The cOmplete His-Tag Purification Resin is an innovative high-capacity IMAC matrix (Immobilized Metal Affinity Chromatography) for convenient single-step purifications of His-tagged proteins from total lysates. Roche′s propriety nickel-chelate chemistry ensures extraordinary compatibility with commonly used reducing agents such as DTT, chelating metalloprotease inhibitors such as EDTA, and a wide range of buffer substances and salt conditions. The wide choice of compatible ingredients allows optimization of buffers for maximum protein stability and solubility. cOmplete His-Tag Purification Columns are fully compatible with standard purification systems such as ÄKTA Systems (Cytiva). Use the buffer conditions best suited to your proteinKeep your protein comfortable and let it, not your purification resin, determine whether you use DTT, EDTA, or other buffer substances. Repeatedly obtain highly pure proteinSingle step purification without resin recharging. Protect your protein from toxic nickelReduce protein oxidation and aggregation caused by resins that leach nickel. Work in a safe and eco-friendly environmentAvoid handling of toxic nickel and completely eliminate disposal costs.

General description

The 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.

Legal Information

cOmplete is a trademark of Roche

Other Notes

For life science research only. Not for use in diagnostic procedures.

Recommended volumetric flow rate: 5 ml column (06 781 535 001): 2.5 to 10 ml/minute1 ml column (06 781 543 001): 0.5 to 2