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| Product Name | His-Tag Protein Purification Resin (Ni-NTA Agarose) |
| Catalog No. | PTR-HMM-0069 |
| Description | A nickel-nitrilotriacetic acid agarose resin for the affinity purification of recombinant proteins bearing a polyhistidine tag. The resin provides high binding capacity and selectivity for His-tagged proteins from bacterial, yeast, insect, and mammalian expression systems. |
| Intended Use | Affinity purification of recombinant His-tagged proteins from clarified cell lysates under native or denaturing conditions for structural, biochemical, and immunological studies. |
| Principle / Technology | Immobilized nickel ions chelated by nitrilotriacetic acid groups on the agarose matrix coordinate with the imidazole side chains of the polyhistidine tag, enabling selective protein capture and subsequent imidazole-gradient elution. |
| Detection Method | Batch or column chromatography with UV absorbance monitoring at 280 nm for protein detection in eluted fractions |
| Sample Type | Clarified bacterial, yeast, insect, or mammalian cell lysates containing overexpressed His-tagged recombinant proteins |
| Performance Range / Specifications | Binding capacity: ≥40 mg of 6×His-tagged protein per milliliter of settled resin; typical purity >90% can be achieved in a single purification step |
| Sensitivity / LOD | Capable of purifying His-tagged proteins present at levels as low as 0.1% of total cellular protein |
| Specificity | High selectivity for His-tagged proteins; non-specific binding of host cell proteins is minimized with appropriate imidazole concentration (10–40 mM) in binding and wash buffers |
| Reaction Conditions / Protocol | Equilibrate column with binding buffer, load clarified lysate, wash with binding buffer containing 20–40 mM imidazole to remove non-specifically bound proteins, elute His-tagged protein with buffer containing 250–500 mM imidazole, regenerate resin with stripping buffer |
| Components / Formulation | Ni-NTA agarose resin as a 50% slurry in 20% ethanol, included buffers: binding buffer (50 mM NaH2PO4, 300 mM NaCl, 10 mM imidazole, pH 8.0), wash buffer (50 mM NaH2PO4, 300 mM NaCl, 20 mM imidazole, pH 8.0), elution buffer (50 mM NaH2PO4, 300 mM NaCl, 250 mM imidazole, pH 8.0) |
| Storage Conditions | 2–8°C; do not freeze the resin slurry; ethanol in the storage solution prevents microbial growth |
| Shelf Life | 24 months from date of manufacture |
| Package Specifications | 10 mL of 50% resin slurry (5 mL settled bed volume) |
| Product Form | 50% agarose bead slurry in 20% ethanol storage solution |
| Quality Control | Each lot tested for His-tagged protein binding capacity using a standardized 6×His-GFP fusion protein; nickel ion leakage tested by ICP-MS; resin stability validated through 10 adsorption-elution-regeneration cycles |
| Key Features | NTA tetradentate chelator provides higher nickel binding stability compared to IDA resins, allowing higher imidazole concentrations in wash buffers; compatible with both gravity-flow and FPLC chromatography systems; reusable for multiple purification cycles |
For research use only, not for clinical use.
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