His-tagged Fusion Protein Purification

We offer two ligands for the purification of His-tagged proteins: nickel (as Ni-NTA) and cobalt. Ni-NTA resins are recommended when maximum protein yield is desired. Cobalt resins are recommended when maximum protein purity is the goal. These magnetic beads and other resins are available in multiple formats to accommodate a variety of needs, from high-throughput screening, batch, pilot, and process purification. Superflow agarose resins have undergone extensive chemical characterization.

  • More formats—magnetic beads, magnetic agarose, loose resin, spin columns and kits, FPLC cartridges, and 96-well filter plates enable fusion protein purification from microgram to kilogram scales.
  • High performance—resins are designed to maximize protein yield and reduce background.
  • Economical—pricing is similar to or better than other suppliers.

Choose a product for His-tagged fusion protein purification

Product type Pierce Ni-NTA Magnetic Beads Pierce Ni-NTA Magnetic Agarose Beads HisPur Ni-NTA Agarose Resin HisPur Ni-NTA Superflow Resin HisPur Cobalt Agarose Resin HisPur Cobalt Superflow Resin
Bead or resin size 1 μm 10–40 μm 45–165 μm 60–160 μm 45–165 μm 60–160 μm
Static binding capacity ≥0.5 mg/mL ≥70 mg/mL ~60 mg/mL ≥60 mg/mL ≥15 mg/mL ≥30 mg/mL
Dynamic binding capacity* N/A N/A 18 mg/mL 20 mg/mL Not determined 20 mg/mL
Maximum linear flow rate* N/A N/A 700 cm/hr 1,200 cm/hr 700 cm/hr 1,200 cm/hr
Support Magnetite-coated polymer Magnetite-embedded 6% beaded agarose 6% beaded agarose Highly crosslinked 6% beaded agarose 6% beaded agarose Highly crosslinked 6% beaded agarose
No. of reuses 0 0 5 25 5 25
Formats available loose beads loose beads loose beads, pre-packed spin columns and kits, chromatography cartridges loose beads loose beads, pre-packed spin columns and kits, chromatography cartridges loose beads
Recommended application scale screening high-throughput batch batch, pilot batch, pilot, process batch, pilot batch, pilot, process
Order product(s) 88831 78605 88221
(loose resin)
90098
(1 mL cartridge)
90099
(5 mL cartridge)
88230
(spin plate)
25214 89964
(loose resin)
90093
(1 mL cartridge)
90094
(5 mL cartridge)
90095
(spin plate)
25228

Additional HisPur Ni-NTA and HisPur Cobalt products

Resin Format Resin per column Column size Collection tube size Cat. No.
Ni-NTA Spin Column 0.2 mL 0.8 mL Microcentrifuge 88224
Ni-NTA Spin Column 1 mL 2 mL 15 mL conical 88225
Ni-NTA Spin Column 3 mL 10 mL 50 mL conical 88226
Ni-NTA Spin Column Kit 0.2 mL 0.8 mL Microcentrifuge 88227
Ni-NTA Spin Column Kit 1 mL 2 mL 15 mL conical 88228
Ni-NTA Spin Column Kit 3 mL 10 mL 50 mL conical 88229
Cobalt Spin Column 0.2 mL 0.8 mL Microcentrifuge 89967
Cobalt Spin Column 1 mL 2 mL 15 mL conical 89968
Cobalt Spin Column 3 mL 10 mL 50 mL conical 89969
Cobalt Spin Column Kit 0.2 mL 0.8 mL Microcentrifuge 90090
Cobalt Spin Column Kit 1 mL 2 mL 15 mL conical 90091
Cobalt Spin Column Kit 3 mL 10 mL 50 mL conical 90092

Featured product data

Comparison of protein yield and purity between Pierce Ni-NTA Magnetic Agarose and competing products from other suppliers. Samples (0.5 mL) of three proteins (6xHis-tagged BirA, 6xHis-tagged red firefly luciferase, or 6xHis-tagged GFP) were diluted with 0.5 mL binding buffer and purified manually with 25 mL settled beads. Respective suppliers’ protocols were followed for their buffer compositions and volumes. Pierce Ni-NTA Magnetic Agarose had the highest yield and similar purity compared to beads from the other suppliers.

Product comparison with Thermo Scienitific HisPur Nickel Superflow Agarose and Qiagen Ni-NTA Superflow resin.

Purity of 6xHis-GFP on nickel resins. Aliquots (8 mL) of an E. coli over-expression lysate corresponding to approx. 16 mg 6xHis-GFP were loaded onto 1 mL columns (diameter = 0.7 cm) of two different Ni-NTA resins: Thermo Scientific HisPur Nickel Superflow Agarose and Qiagen™ Ni-NTA Superflow (Cat. No. 30410). Columns were washed with 10 mL (10 column volumes) of wash buffer containing 30 mM imidazole. Bound protein was eluted with 10 mL of elution buffer containing 300 mM imidazole. Fractions were collected at 1 mL intervals. Aliquots of load lysate (L), flow-through (FT), wash (W), and eluate (1 to 10) fractions were separated by SDS-PAGE and stained with Thermo Scientific Imperial Protein Stain (Cat. No. 24615). Purity of 6xHis-GFP across all elution fractions in each gel was determined by densitometry with Thermo Scientific myImageAnalysis Software (Cat. No. 62237). Read the full application note: Performance Characterization of HisPur Nickel Superflow Agarose.