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Protein Expression & Purification

Products >  Protein_Expression_and_Purification >  His-Tagged_Protein_Purification >  Nickel_Resin_Batch_FPLC

His-Tagged Recombinant Protein Purification—His60 Ni Resin

His60 Ni Superflow Resin is a high-capacity Ni-IDA resin for the one-step purification of recombinant his-tagged proteins from bacterial, mammalian, and baculovirus-infected cells. The modified Ni-IDA chemistry makes this resin one of the best on the market in terms of capacity, metal ion leakage, and purity, when compared to other Ni-based resins such as Ni-NTA or other Ni-IDA resins. See how His60 outperforms Ni-NTA here.

The Recombinant His-Tagged Protein Purification Principle

His60 Ni Resin is compatible with batch/gravity flow applications, as well as with the major automated liquid chromatography systems and manual syringe processing.The His60 Ni Superflow purification resin has low metal ion leakage and the highest binding capacity to date (outperforming Ni-NTA and other resins), as well as low metal ion leakage. The resin enables fast, easy, and reproducible chromatographic separations and can be regenerated for multiple uses. For additional information, please refer to the User Manual for Recombinant His-Tagged Protein Purification.

The Recombinant His-Tagged Protein Purification Procedure

The purification of recombinant his-tagged proteins consists of 4 steps: cell lysis/extraction, binding, washing, and elution. His60 Ni allows one-step protein purification under either native or denaturing conditions. The resin is compatible with multiple denaturants and detergents—refer to the User Manual for a complete reagent compatibility table. Purified recombinant his-tagged proteins can be eluted by the addition of 300 mM imidazole or by a reduction in pH.

  At-A-Glance   Documents   Images & Data   Resources


  • Up to 60 mg/ml of recombinant his-tagged protein
  • Low metal ion leakage in eluted his-tagged protein
  • For purification under native or denaturing conditions
  • Use for batch/gravity flow and automated FPLC purification


Purified recombinant his-tagged proteins can be used for:

  • Protein labeling
  • Immunization to produce antibodies
  • Animal studies
  • Other applications

We Also Recommend

  • His-Tag Detection Antibodies
    Use highly sensitive antibodies to detect his-tagged recombinant proteins in Western blot, ELISA, and immunocytohistochemical assays
  • ProteoGuard Protease Inhibitor Cocktail 
    Use this protease inhibitor cocktail to suppress proteolysis in all your cell lysates. We recommend using ProteoGuard with all our protein purification products

Reagents Compatible with His60 Ni Superflow Resin

Reagent Acceptable Concentration
Beta-mercaptoethanola 20 mM (with caution)
CaCl2 5 mM
CHAPSb,c 1% (with caution)
DTT (dithiothreitol)d 1 mM (with caution)
Ethanole 20%
Glycerol 20%
Guanidine hydrochloridef 6 mM
HEPESg Up to 100 mM (with caution)
Histidineh 20 mM to inhibit nonspecific binding.
Up to 100 mM to elute his-tagged proteins.
MgCl2 4 M
MOPSg Up to 100 mM (with caution)
NP-40b,i 2%
SDSb,c 1% (with caution)
Sodium acetate Up to 100 mM (with caution)
Sodium phosphate Up to 50 mM
Trisj Up to 50 mM (with caution)
Triton-X 100b,i 1%
Tween 20 2%
Ureaf 8 M

a Use the resin immediately after equilibrating with buffers containing beta-mercaptoethanol. Otherwise, a slight change in color (yellowing of the resin) will occur. Do not store the resin in buffers containing beta-mercaptoethanol.
b Detergents cannot be easily removed by buffer exchange.
c Ionic detergents like CHAPS (3-[(3-Cholamidopropyl)-dimethylammonio]-1-propane-sulfonate), SDS (sodium dodecyl sulfate), and sarkosyl are compatible up to 1%. However, due to their charged nature, you should anticipate interference with binding, even at low concentrations.
Since DTT is a reducing agent, low concentrations will reduce the metal ions in His60 Ni Superflow resin. Although enough of these ions may remain unaffected to allow protein purification, please use it with caution. Do at least 20 column volumes of washes, preferably with low concentrations of imidazole (40 mM) to wash out any reduced metal ions.
e Ethanol may precipitate proteins, causing low yields and column clogging.
f With high concentrations, protein unfolding generally takes place. Protein refolding on-column (or after elution) is protein-dependent.
g Amine groups that are present in these buffers can interact with Ni2+ ions, diminishing the resin’s binding capacity.
h Binds to His60 Ni and competes with histidine residues in the his tag.
i Has high absorbance at 280 nm.
j Tris coordinates weakly with metal ions, causing a decrease in capacity.

Reagents Incompatible with His60 Ni Superflow Resin

These reagents are incompatible at any concentration:

  • Arginine, glycine, and glutamine

  • EDTA (ethylenediaminetetraacetic acid), EGTA (ethylene glycol tetraacetic acid), and PEI
    NOTE: Using these chelating agents will strip metal ions from the resin, resulting in protein elution and a resin color change.

  • DTE (dithioerythritol)

Additional Product Information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.


Cat. # Product Contents Size Price Units Select
635665 His60 Ni Buffer Set 20 Purifications $198.00 *
635659 His60 Ni Superflow Resin 10 mL $74.40 *
635660 His60 Ni Superflow Resin 25 mL $180.00 *
635661 His60 Ni Superflow Resin 4 x 25 mL $690.00 *
635662 His60 Ni Superflow Resin 250 mL $1,567.50 *
635663 His60 Ni Superflow Resin 2 x 250 mL $2,736.00 *
635664 His60 Ni Superflow Resin 4 x 250 mL $4,860.00 *
635677 His60 Ni Superflow Resin & Buffer Set Bundle 20 Purifications Call *
635676 His60 Ni Superflow Resin & xTractor™ Buffer Bundle 20 Purifications Call *
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