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PureCube Compact Cartridge Co-NTA
The term Co-NTA refers to a copper Ion that has been coupled Nitrilotriacetic acid. Co-NTA can then be coupled to agarose or magnetic beads for IMAC (Immobilized Metal Chelate Affinity Chromatography). This is a purification method to obtain functional His-tagged protein. Cube Biotech offers multiple different Co-NTA based products for protein purification. FPLC based protein purification of His-tagged proteins is a common practice. Therefore we pre-filled our PureCube COMPACT Cartridges / Columns with our PureCube Co-NTA agarose resin. The columns are suited for most FPLC devices. However please check the corresponding "Features" table to confirm compability with your machinery. In case of no compability we offer cartridges / columns cartridges / columns of different formats pre-filled with Co-NTA. Co-NTA agarose COMPACT cartridges with 100 µm diameter agarose beads are to go-to option if higher flow rates are needed.
1. Affinity Resin
PureCube Co-NTA Agarose was developed for the affinity purification of proteins carrying a polyhistidine tag. This affinity chromatography matrix is based on BioWorks Workbeads, consisting of 7.5% crosslinked agarose. The material is highly porous to allow for optimal protein interaction. Cross-linked agarose is also physically very stable, making it suitable for purification processes under low pressure with variable flow rates. Our agarose is very homogeneous in size with a medium particle diameter of 40 µm, yielding a high degree of reproducibility between individual purification runs.
An NTA ligand is coupled to the agarose resin and carefully loaded with cobalt ions to obtain a matrix with high specificity and binding capacity for histidine residues. The metal ion capacity is about 15 µeqv Co2+/mL. Other possible metal ions are Ni2+, Zn2+, Fe3+, Al3+, resulting in different affinities, e.g., for zinc-finger proteins or phosphorylated proteins. If required, the nickel ions can be removed from the agarose matrix using five wash steps with 100 mM EDTA, and the matrix can be recharged with a different metal ion. Alternatively, please contact us for unloaded NTA agarose. NTA matrices charged with other metals are available upon request.
2. Protein Binding Capacity
PureCube Compact Cartridges Co-NTA have a binding capacity of up to 20 mg/mL as determined by purification of 6xHis-tagged GFP protein from E.coli cleared lysates, and quantified via spectrophotometry. It should be considered that the dynamic binding capacity strongly correlates with flow rate and other parameters such as protein size and buffer conditions. It is recommended to use the lowest flow rate possible to achieve highest binding capacity.
3. Compatibility
For cleaning purposes, PureCube Co-NTA Agarose is very stable and can resist the following conditions in most situations: All commonly used aqueous buffers, pH 2-14, 100% methanol, 100% ethanol, 8 M urea, 6 M guanidinium hydrochloride, 30% (v/v) acetonitrile, and up to 10 mM DTT.
Features - Regarding Purification capabilities.
Type: | Cartridge |
Ligand: | NTA |
Coupled Ion: | Co2+ |
Usage | Specific binding and purification of 6x His-tagged proteins |
Specifity | Affinity to His-tagged proteins |
Binding capacity | >30 mg/mL |
Bead Ligand | Co-NTA |
Bead size | 40 μm(Agarose) |
Chelator stability | Stable in buffer containing 10 mM DTT and 1 mM EDTA |
ph Stability | 2-14 |
Required equipment |
· -Lysis Buffer · -Wash Buffer · -Elution Buffer · -Ice bath · -Refrigerated centrifuge for 50 mL tube (min 10,000 x g) · -50 mL centrifuge tube · -Micropipettor and Micropipetting tips · -Disposable gravity flow columns with capped bottom outlet, 2 ml · - pH meter · -End-over-end shaker · -SDS-PAGE buffers, reagents and equipment Optional: Western Blot reagents and equipment |
Order number | Products | Bead size | Protein Binding Capacity | Shipment Temperature | Storage temperature | Usage | Price(yuan) |
31602 | PureCube Compact Cartridge Co-NTA(1×1ml) | 40 | 30mg | Ambient temperature | 4° C | Use for highly specific binding and purification of 6x His-tagged proteins; Stable in buffer containing 10 mM DTT and 1 mM EDTA; pH Stability: 2-14; Max. Flow Rate: 6 mL/min | 593 |
31604 | PureCube Compact Cartridge Co-NTA(5×1ml) | 40 | 30mg | Ambient temperature | 4° C | 2649 | |
31606 | PureCube Compact Cartridge Co-NTA(1×5ml) | 40 | 150mg | Ambient temperature | 4° C | 1728 | |
31608 | PureCube Compact Cartridge Co-NTA(5×5ml) | 40 | 150mg | Ambient temperature | 4° C | 7698 |
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