- Quick and easy separation of his-tagged proteins
- Selective His60 Ni chemistry for increased purity
- Eluted samples ideal for small volume applications
His60 Ni Magnetic Beads provide simple, effective separation of recombinant his-tagged proteins for a wide range of applications, including:
- Microscale purification of his-tagged proteins
- Studying protein structure and function
- Preparing proteins for x-ray crystallography
- Performing assays that detect protein-protein and protein-DNA interactions
- Immunization to raise antibodies against a protein of interest
- Screening for protein expression
- Optimizing purification conditions for scale-up with His60 Ni Resin
His60 Ni Magnetic Beads can also be used to immobilize 6xHis-tagged proteins for affinity chromatography, in order to:
- Analyze protein-protein and protein-nucleic acid interactions
- Purify untagged subunits or nucleic acids that interact with the immobilized protein
- Perform antibody purification
- Study interactions between ligands and receptors
- Choice of native or denaturing conditions to purify proteins
- Directed presentation of 6xHis-tagged proteins increases reproducibility and signal-to-noise ratios
- Varying the amount of beads per well allows for a wide range of binding capacities
- A powerful tool for analyzing interactions between biomolecules
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)|
|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)|
|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.
d 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.