Ni-NTA Resin

The Ni-NTA Resin in a high binding, high capacity nickel charged IMAC resin that purifies recombinant proteins with the polyhistidine (6XHis) sequence.  The resin is 6% cross-linked agarose for excellent structural integrity with a high capacity of 20-40μmoles Ni²⁺/ml resin that results in high protein binding capacity of >50mg protein per ml resin.  In fact, we have demonstrated binding of >100mg/ml resin of a 50kDa 6XHis tagged protein. Available as ready-to-use resin in 5 convenient sizes (10, 25, 100, 500 and 2 x 500ml) or supplied prepacked in various spin column formats (0.2, 1 and 3ml) and spin plates.

Features

• High capacity: >50mg/ml
• Ligand density: 20-40μmoles Ni²⁺ /ml resin
• Bead Structure: 6% cross-linked agarose
• Uses nitrilotriacetic acid (NTA), a tetradenate chelating ligand
• For the purification of 6x His proteins

Immobilized Metal Ion Affinity Chromatography (IMAC), developed by Porath (1975), is based on the interaction of certain protein residues (histidines, cysteines, and to some extent tryptophans) with cations of transition metals. The Ni-NTA Resin is specifically designed for the purification of recombinant proteins fused to the polyhistidine (6XHis) tag expressed in bacteria, insects, and mammalian cells.  The resin is high affinity and selectivity for recombinant fusion proteins that are tagged with six tandem histidine residues. The Ni-NTA resin can be used to purify 6X His tagged proteins under native and denaturing conditions.  Proteins bound to the resin can be eluted with low pH buffer or competition with imidazole or histidine. The Ni-NTA resin uses nitrilotriacetic acid (NTA), a tetradenate chelating ligand, in a highly cross-linked 6% agarose matrix.  The NTA binds Ni²⁺ ions by four coordination sites.

The spin columns are supplied with a resin bed volume of 0.2, 1 and 3ml with total column volumes of 1, 8 and 22ml respectively. Columns can be used as a spin format of gravity flow columns. Also available in 96-well spin plate formats for processing up to 96 samples.

Immobilized IDA Nickel,  Copper, Cobalt  and Zinc  Chelating Resins and Cobalt NTA Resin are also available.  Cobalt has the highest selectivity followed by Zinc, Nickel then Copper, but has the lowest binding capacity.  Copper has the highest binding capacity, followed by Nickel then Zinc.

Specific binding/wash and elution buffers are available.

Our HOOK™ 6X His Protein Purification kits include everything required for purification of 6X His tagged proteins from yeast or bacteria, including lysis buffers, lytic enzymes, resin, columns and binding, wash and elution buffers. Each kit is available with nickel or cobalt chelating resins. Kits available are:

• HOOK™ 6X His Protein Purification (Bacteria): Isolation from bacteria; optimized to yield up to 10mg/250ml culture of soluble 6X His tagged protein.
• HOOK™ 6X His Protein Spin Purification (Bacteria): Isolation from bacteria;optimized to yield ~1mg/50ml culture of soluble 6X His tagged protein
• HOOK™ 6X His Protein Purification (Yeast): Isolation from yeast; optimized to yield up to 10mg of soluble 6X His tagged protein.
• HOOK™ 6X His Protein Spin Purification (Yeast): Isolation from yeast; optimized to yield up to 1mg of soluble 6X His tagged protein.