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 6 x histidine (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 tand em 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.
This resin has a capacity of 20-40μmoles Ni2+/ml resin. The protein binding capacity is >50mg protein per ml resin. We have demonstrated binding of >100mg of a 50kDa 6XHis tagged proteins to a ml of resin.
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 new HOOK™ 6X His Protein Purification kits are now available and 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. Click here to request bulk or custom sizes
- Uses nitrilotriacetic acid (NTA), a tetradenate chelating ligand
- For the purification of 6x His proteins
- High capacity: >50mg/ml
- Ligand density: 20-40μmoles Ni2+ /ml resin
- Bead Structure: 6% cross-linked agarose
- Affinity purification of proteins with a 6x His tag.