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Products > Protein_Expression_and_Purification > His-Tagged_Protein_Purification > Nickel_Columns-FPLC

His60 Ni Superflow Cartridges Using FPLC

His 60 Ni Superflow, the highest-capacity resin to date that provides great purity, is now available in prefilled 1 ml and 5 ml cartridges for his-tagged protein purification on any LC system, such as ÄKTA or FPLC, or for manual purification using a syringe. The resin enables fast, easy, and reproducible chromatographic separations and can be regenerated for multiple uses.

At-A-Glance

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Features

Up to 60 mg/ml of recombinant his-tagged protein (1 ml cartridge)
Up to 300 mg/ml of recombinant his-tagged protein (5 ml cartridge)
Low metal ion leakage in eluted his-tagged protein
For purification under native or denaturing conditions
Fast and reproducible processing on any LC system, including ÄKTA

Applications

Purified recombinant his-tagged proteins can be used for:

Protein labeling
Immunization to produce antibodies
Animal studies
Other applications

His60 Ni Superflow Cartridge Specifications

	1 ml Cartridge	5 ml Cartridge
Support	Superflow 6 (cross-linked agarose)
Bead Diameter	60–160 µm
Column Dimensions (mm i.d.)	0.7 cm x 2.5 cm	1.6 cm x 2.5 cm
Maximum Pressure¹	5 bar, 0.5 MPa
Typical Back Pressure	1.0 bar, 0.1 MPa (1 ml/min)	2.0 bar, 0.2 MPa (5 ml/min)
Recommended Flow Rate	1 ml/min (156 cm/hr)	5 ml/min (149 cm/hr)
Maximum Flow Rate²	10 ml/min (1,559 cm/hr)	40 ml/min (1,193 cm/hr)
pH Stability Short Term (<2 hr)	2–14
pH Stability Long Term (>2 hr)	3–14
Binding Capacity³	60 mg of AcGFPuv	300 mg of AcGFPuv
System Compatibity⁴	Automated chromatography systems (e.g. ÄKTA, FPLC, etc.), peristaltic pump or syringe
Cartridge Body Material	Polypropylene
Connectors	1/16"(inlet); 1/16" (outlet)

¹ The maximum pressure that can be used with the Superflow matrix itself is 10 bars. However, stability of the cartridges is only guaranteed up to 5 bars.
² High flow rates may lead to reduced recovery of 6xHis-tagged protein.
³ Binding capacity may vary from protein to protein.
⁴ Adaptors may be necessary.

Additional Products

His60 Ni resin (a nickel-based resin) is sold separately as:

TALON resin (a cobalt-based resin) is sold separately as:

Detect his-tagged proteins with his tag antibodies

Reagents Compatible with His60 Ni Superflow Resin

Reagent	Acceptable Concentration
Beta-mercaptoethanol^a	20 mM (with caution)
CaCl₂	5 mM
CHAPS^b,c	1% (with caution)
DTT (dithiothreitol)^d	1 mM (with caution)
Ethanol^e	20%
Glycerol	20%
Guanidine hydrochloride^f	6 mM
HEPES^g	Up to 100 mM (with caution)
Histidine^h	20 mM to inhibit nonspecific binding. Up to 100 mM to elute his-tagged proteins.
MgCl₂	4 M
MOPS^g	Up to 100 mM (with caution)
NP-40^b,i	2%
SDS^b,c	1% (with caution)
Sodium acetate	Up to 100 mM (with caution)
Sodium phosphate	Up to 50 mM
Tris^j	Up to 50 mM (with caution)
Triton-X 100^b,i	1%
Tween 20	2%
Urea^f	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.
^dSince 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 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.