Description
GoldBio's Nickel NTA HTC Agarose Beads are high-capacity immobilized metal affinity chromatography (IMAC) resins designed for His-tagged protein purification in applications that need high flow rates (up to 300 kPa) and high operating pressures.
The highly crosslinked agarose matrix is compatible with FPLC and HPLC systems and withstands pressures up to 300 kPa, making it well suited for laboratory-scale purification, process development, and protein purification scale-up studies.
The resin provides a binding capacity of approximately 60 mg of His-tagged protein per mL of gel and supports purification under both native and denaturing conditions.
Nickel NTA HTC Agarose Beads are commonly used to purify recombinant proteins expressed in bacterial, yeast, insect, and mammalian expression systems. Their mechanical strength and consistent flow properties make them a good choice for protein purification protocols that require rapid processing, controlled flow, and reproducible chromatographic performance. The resin is supplied as a ready-to-use suspension and may be autoclaved according to the validated product handling instructions.
Purification is based on immobilized metal affinity chromatography (IMAC). Nitrilotriacetic acid (NTA) groups are covalently attached to the highly crosslinked agarose matrix and charged with nickel ions (Ni²⁺). The immobilized nickel selectively binds polyhistidine affinity tags engineered onto recombinant proteins, allowing target proteins to be separated from complex lysates.
Compared with IDA-based resins, NTA chemistry generally provides greater nickel ion stability and reduced susceptibility to metal leaching under compatible purification conditions. Target proteins are typically recovered by competitive elution with imidazole using standard His-tag purification protocols.
HTC beads are high-throughput beads designed to meet the demand for industrial process separation. Their rigidity and mechanical resistance permit high flow rates, with good resolution in a minimum time frame, making these beads ideal for process-scale use. HTC beads are highly crosslinked 4% and 6% agarose matrices and have excellent physical and chromatographic qualities. They are autoclavable. These beads are an ideal support for the immobilization of ligands for Affinity Chromatography and base media support for producing IEX (Ion Exchange) and Hydrophobic interaction chromatography resins
Suitability: Nickel-NTA HTC agarose resin is suitable for use with FPLC or HPLC, and can withstand pressures up to 300 kPa.
Binding Capacity: ≥60 mg/ml gel
Note: GoldBio recommends using Nickel NTA resins in the presence of elevated levels of reducing agents in order to ensure optimal purification of your target protein. Nickel IDA resins may show some discoloration in reaction with low levels of reducing agents but are still functional at low levels of reducing agents (≤5mM DTT).
Common Applications
(Click each for more information)
Purification of His-Tagged Recombinant Proteins
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Purpose: To selectively isolate recombinant proteins containing engineered polyhistidine (His) tags from complex biological samples.
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How It Works: Histidine residues within the affinity tag coordinate with nickel ions immobilized on the NTA agarose matrix, enabling selective binding while unbound proteins are removed during washing. Target proteins are typically eluted using imidazole.
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Applications: Recombinant protein purification, enzyme isolation, antibody fragment purification, protein production, and routine laboratory protein purification.
Bornhorst, J. A., & Falke, J. J. (2000). Purification of proteins using polyhistidine affinity tags. Methods in Enzymology, 326, 245–254. https://doi.org/10.1016/S0076-6879(00)26058-8
High-Pressure FPLC and HPLC Protein Purification
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Purpose: To purify His-tagged proteins using chromatography systems that operate at elevated flow rates and pressures.
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How It Works: The highly crosslinked HTC agarose matrix resists compression under increased operating pressures, maintaining consistent flow characteristics and chromatographic performance during automated purification.
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Applications: FPLC purification, HPLC-based affinity chromatography, automated protein purification, and process development.
Block, H., Maertens, B., Spriestersbach, A., et al. (2009). Immobilized-metal affinity chromatography (IMAC): A review. Methods in Enzymology, 463, 439–473. https://doi.org/10.1016/S0076-6879(09)63027-5
Scalable Recombinant Protein Purification
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Purpose: To develop and scale His-tagged protein purification workflows that require increased throughput and mechanically stable chromatography media.
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How It Works: The mechanically robust HTC matrix supports increased flow rates and larger sample loads while maintaining controlled chromatographic operation within the stated pressure limit.
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Applications: Process development, purification scale-up, pilot studies, recombinant protein production, and optimization of chromatography conditions.
Arnau, J., Lauritzen, C., Petersen, G. E., & Pedersen, J. (2006). Current strategies for the use of affinity tags and tag removal for the purification of recombinant proteins. Protein Expression and Purification, 48(1), 1–13. https://doi.org/10.1016/j.pep.2005.12.002
Protein Purification Under Native and Denaturing Conditions
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Purpose: To recover soluble recombinant proteins or purify His-tagged proteins from insoluble inclusion bodies.
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How It Works: Nickel NTA affinity can be used under native conditions or in compatible denaturing buffers containing urea or guanidine hydrochloride, allowing purification strategies to be adapted to soluble proteins and proteins recovered from inclusion bodies.
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Applications: Purification of soluble proteins, inclusion body recovery, protein refolding studies, and recombinant protein production.
Block, H., Maertens, B., Spriestersbach, A., et al. (2009). Immobilized-metal affinity chromatography (IMAC): A review. Methods in Enzymology, 463, 439–473. https://doi.org/10.1016/S0076-6879(09)63027-5
Structural Biology and Protein Characterization
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Purpose: To obtain highly purified recombinant proteins suitable for downstream structural and biochemical analyses.
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How It Works: Selective affinity purification enriches the His-tagged target and reduces host-cell contaminants before polishing steps or downstream structural and functional analyses.
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Applications: Protein crystallography, cryo-EM sample preparation, enzyme characterization, biophysical analysis, and protein-protein interaction studies.
Terpe, K. (2003). Overview of tag protein fusions: From molecular and biochemical fundamentals to commercial systems. Applied Microbiology and Biotechnology, 60(5), 523–533. https://doi.org/10.1007/s00253-002-1158-6
Key Benefits
· High-Pressure Compatibility: The highly crosslinked HTC agarose matrix withstands operating pressures up to 300 kPa, supporting use with compatible FPLC and HPLC systems.
· Scalable Protein Purification: Supports routine laboratory purification, process-development studies, and scale-up of compatible His-tag protein purification.
· High Specificity with Low Metal Ion Leaching: he tetradentate NTA ligand generally provides greater nickel ion stability than IDA-based resins while supporting selective binding of His-tagged proteins.
· Compatible with Native and Denaturing Conditions: Enables purification of soluble recombinant proteins as well as recovery of His-tagged proteins from inclusion bodies using standard denaturing purification protocols.
· High Mechanical Stability: The rigid, highly crosslinked agarose matrix resists bead compression at elevated flow rates and supports controlled chromatographic operation within the stated pressure limit.
Storage/Handling:
Store at 4°C. Do not freeze.
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