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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 High-Capacity Ni-IMAC MagBeads, EDTA Compatible | Pierce Ni-NTA Magnetic Agarose Beads | Pierce Ni-NTA Magnetic Beads |
|---|---|---|---|
| Bead or resin size | 100 µm | 10–40 μm | 1 μm |
| Static binding capacity | >80 mg/ml | ≥70 mg/mL | ≥0.5 mg/mL |
| Dynamic binding capacity* | >60 mg/ml | N/A | N/A |
| Maximum linear flow rate* | 300 cm/hr (150 cm/hr recommended) | N/A | N/A |
| Support | 6% crosslinked agarose | Magnetite-embedded 6% beaded agarose | Magnetite-coated polymer |
| No. of reuses | Up to 10 | 0 | 0 |
| Formats available | Loose beads | Loose beads | Loose beads |
| Recommended application scale | Scalable | High-throughput batch | Screening |
| Cat. No. | A50588 (1 mL) A50589 (5 mL) A50590 (25 mL) A50591 (100 mL) | 78605 | 88831 |
| Product type | HisPur Ni-NTA Agarose Resin | HisPur Ni-NTA Superflow Resin | Pierce High-Capacity Ni-IMAC Resin, EDTA Compatible | HisPur Cobalt Agarose Resin | HisPur Cobalt Superflow Resin |
|---|---|---|---|---|---|
| Bead or resin size | 45–165 μm | 60–160 μm | 100 µm | 45–165 μm | 60–160 μm |
| Static binding capacity | ~60 mg/mL | ≥60 mg/mL | >80 mg/mL | ≥15 mg/mL | ≥30 mg/mL |
| Dynamic binding capacity* | 18 mg/mL | 20 mg/mL | >60 mg/mL | Not determined | 20 mg/mL |
| Maximum linear flow rate* | 700 cm/hr | 1,200 cm/hr | 300 cm/hr | 700 cm/hr | 1,200 cm/hr |
| Support | 6% beaded agarose | Highly crosslinked 6% beaded agarose | Highly cross-linked 6% agarose | 6% beaded agarose | Highly crosslinked 6% beaded agarose |
| No. of reuses | 5 | 25 | 10 | 5 | 25 |
| Formats available | Loose beads, pre-packed spin columns and kits, chromatography cartridges | Loose beads | Loose beads | Loose beads, pre-packed spin columns and kits, chromatography cartridges | Loose beads |
| Recommended application scale | Batch, pilot | Batch, pilot, process | Batch, pilot | Batch, pilot | Batch, pilot, process |
| Cat. No. | 88221 (loose resin) 90098 (1 mL cartridge) 90099 (5 mL cartridge) 88230 (spin plate) | 25214 | A50584 (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 type | Pierce High-Capacity Ni-IMAC MagBeads, EDTA Compatible | Pierce Ni-NTA Magnetic Agarose Beads | Pierce Ni-NTA Magnetic Beads |
|---|---|---|---|
| Bead or resin size | 100 µm | 10–40 μm | 1 μm |
| Static binding capacity | >80 mg/ml | ≥70 mg/mL | ≥0.5 mg/mL |
| Dynamic binding capacity* | >60 mg/ml | N/A | N/A |
| Maximum linear flow rate* | 300 cm/hr (150 cm/hr recommended) | N/A | N/A |
| Support | 6% crosslinked agarose | Magnetite-embedded 6% beaded agarose | Magnetite-coated polymer |
| No. of reuses | Up to 10 | 0 | 0 |
| Formats available | Loose beads | Loose beads | Loose beads |
| Recommended application scale | Scalable | High-throughput batch | Screening |
| Cat. No. | A50588 (1 mL) A50589 (5 mL) A50590 (25 mL) A50591 (100 mL) | 78605 | 88831 |
| Product type | HisPur Ni-NTA Agarose Resin | HisPur Ni-NTA Superflow Resin | Pierce High-Capacity Ni-IMAC Resin, EDTA Compatible | HisPur Cobalt Agarose Resin | HisPur Cobalt Superflow Resin |
|---|---|---|---|---|---|
| Bead or resin size | 45–165 μm | 60–160 μm | 100 µm | 45–165 μm | 60–160 μm |
| Static binding capacity | ~60 mg/mL | ≥60 mg/mL | >80 mg/mL | ≥15 mg/mL | ≥30 mg/mL |
| Dynamic binding capacity* | 18 mg/mL | 20 mg/mL | >60 mg/mL | Not determined | 20 mg/mL |
| Maximum linear flow rate* | 700 cm/hr | 1,200 cm/hr | 300 cm/hr | 700 cm/hr | 1,200 cm/hr |
| Support | 6% beaded agarose | Highly crosslinked 6% beaded agarose | Highly cross-linked 6% agarose | 6% beaded agarose | Highly crosslinked 6% beaded agarose |
| No. of reuses | 5 | 25 | 10 | 5 | 25 |
| Formats available | Loose beads, pre-packed spin columns and kits, chromatography cartridges | Loose beads | Loose beads | Loose beads, pre-packed spin columns and kits, chromatography cartridges | Loose beads |
| Recommended application scale | Batch, pilot | Batch, pilot, process | Batch, pilot | Batch, pilot | Batch, pilot, process |
| Cat. No. | 88221 (loose resin) 90098 (1 mL cartridge) 90099 (5 mL cartridge) 88230 (spin plate) | 25214 | A50584 (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
| 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.
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
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