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 typePierce High-Capacity Ni-IMAC MagBeads, EDTA CompatiblePierce Ni-NTA Magnetic Agarose BeadsPierce Ni-NTA Magnetic Beads
Bead or resin size100 µm10–40 μm1 μm
Static binding capacity>80 mg/ml≥70 mg/mL≥0.5 mg/mL
Dynamic binding capacity*>60 mg/ml N/AN/A
Maximum linear flow rate*300 cm/hr (150 cm/hr recommended)N/AN/A
Support6% crosslinked agaroseMagnetite-embedded 6% beaded agaroseMagnetite-coated polymer
No. of reusesUp to 1000
Formats availableLoose beadsLoose beadsLoose beads
Recommended application scaleScalableHigh-throughput batchScreening
Cat. No.A50588 (1 mL)
A50589 (5 mL)
A50590 (25 mL)
A50591 (100 mL)
7860588831
Product typeHisPur Ni-NTA Agarose ResinHisPur Ni-NTA Superflow ResinPierce High-Capacity
Ni-IMAC Resin, EDTA Compatible
HisPur Cobalt Agarose ResinHisPur Cobalt Superflow Resin
Bead or resin size45–165 μm60–160 μm100 µm45–165 μm60–160 μm
Static binding capacity~60 mg/mL≥60 mg/mL>80 mg/mL≥15 mg/mL≥30 mg/mL
Dynamic binding capacity*18 mg/mL20 mg/mL>60 mg/mLNot determined20 mg/mL
Maximum linear flow rate*700 cm/hr1,200 cm/hr300 cm/hr700 cm/hr1,200 cm/hr
Support6% beaded agaroseHighly crosslinked 6% beaded agaroseHighly cross-linked 6% agarose6% beaded agaroseHighly crosslinked 6% beaded agarose
No. of reuses52510525
Formats availableLoose beads, pre-packed spin columns and kits, chromatography cartridgesLoose beadsLoose beadsLoose beads, pre-packed spin columns and kits, chromatography cartridgesLoose beads
Recommended application scaleBatch, pilotBatch, pilot, processBatch, pilotBatch, pilotBatch, pilot, process
Cat. No.88221
(loose resin)
90098
(1 mL cartridge)
90099
(5 mL cartridge)
88230
(spin plate)
25214A50584
(20 mL)
A50585
(100 mL)
A50586
(500 mL)
A50587
(1000 mL)
89964
(loose resin)
90093
(1 mL cartridge)
90094
(5 mL cartridge)
90095
(spin plate)
25228
Product typePierce High-Capacity Ni-IMAC MagBeads, EDTA CompatiblePierce Ni-NTA Magnetic Agarose BeadsPierce Ni-NTA Magnetic Beads
Bead or resin size100 µm10–40 μm1 μm
Static binding capacity>80 mg/ml≥70 mg/mL≥0.5 mg/mL
Dynamic binding capacity*>60 mg/ml N/AN/A
Maximum linear flow rate*300 cm/hr (150 cm/hr recommended)N/AN/A
Support6% crosslinked agaroseMagnetite-embedded 6% beaded agaroseMagnetite-coated polymer
No. of reusesUp to 1000
Formats availableLoose beadsLoose beadsLoose beads
Recommended application scaleScalableHigh-throughput batchScreening
Cat. No.A50588 (1 mL)
A50589 (5 mL)
A50590 (25 mL)
A50591 (100 mL)
7860588831
Product typeHisPur Ni-NTA Agarose ResinHisPur Ni-NTA Superflow ResinPierce High-Capacity
Ni-IMAC Resin, EDTA Compatible
HisPur Cobalt Agarose ResinHisPur Cobalt Superflow Resin
Bead or resin size45–165 μm60–160 μm100 µm45–165 μm60–160 μm
Static binding capacity~60 mg/mL≥60 mg/mL>80 mg/mL≥15 mg/mL≥30 mg/mL
Dynamic binding capacity*18 mg/mL20 mg/mL>60 mg/mLNot determined20 mg/mL
Maximum linear flow rate*700 cm/hr1,200 cm/hr300 cm/hr700 cm/hr1,200 cm/hr
Support6% beaded agaroseHighly crosslinked 6% beaded agaroseHighly cross-linked 6% agarose6% beaded agaroseHighly crosslinked 6% beaded agarose
No. of reuses52510525
Formats availableLoose beads, pre-packed spin columns and kits, chromatography cartridgesLoose beadsLoose beadsLoose beads, pre-packed spin columns and kits, chromatography cartridgesLoose beads
Recommended application scaleBatch, pilotBatch, pilot, processBatch, pilotBatch, pilotBatch, pilot, process
Cat. No.88221
(loose resin)
90098
(1 mL cartridge)
90099
(5 mL cartridge)
88230
(spin plate)
25214A50584
(20 mL)
A50585
(100 mL)
A50586
(500 mL)
A50587
(1000 mL)
89964
(loose resin)
90093
(1 mL cartridge)
90094
(5 mL cartridge)
90095
(spin plate)
25228


Additional HisPur Ni-NTA and HisPur Cobalt products

ResinFormatResin per columnColumn sizeCollection tube sizeCat. No.
Ni-NTASpin Column0.2 mL0.8 mLMicrocentrifuge88224
Ni-NTASpin Column1 mL2 mL15 mL conical88225
Ni-NTASpin Column3 mL10 mL50 mL conical88226
Ni-NTASpin Column Kit0.2 mL0.8 mLMicrocentrifuge88227
Ni-NTASpin Column Kit1 mL2 mL15 mL conical88228
Ni-NTASpin Column Kit3 mL10 mL50 mL conical88229
CobaltSpin Column0.2 mL0.8 mLMicrocentrifuge89967
CobaltSpin Column1 mL2 mL15 mL conical89968
CobaltSpin Column3 mL10 mL50 mL conical89969
CobaltSpin Column Kit0.2 mL0.8 mLMicrocentrifuge90090
CobaltSpin Column Kit1 mL2 mL15 mL conical90091
CobaltSpin Column Kit3 mL10 mL50 mL conical90092


Featured product data

Comparison of protein yield and purity between Pierce Ni-NTA Magnetic Agarose and competing products from other suppliers.

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