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| Product Name | Imidazole Buffer, 2 M, pH 7.0, for His-Tag Protein Purification |
| Catalog No. | ATR-B-YJL-0196 |
| Description | 2 M imidazole buffer solution, pH 7.0, specifically formulated for immobilized metal affinity chromatography (IMAC) purification of recombinant His-tagged proteins. Imidazole competes with the polyhistidine tag for binding to Ni²⁺ or Co²⁺ charged resin, enabling step-gradient or linear-gradient elution of His-tagged proteins. This high-concentration stock simplifies preparation of elution buffers at typical working concentrations (20-500 mM imidazole). |
| Intended Use | Elution of His-tagged proteins from Ni-NTA, Co-NTA, and other IMAC resins; competitive elution in metal affinity chromatography; preparation of imidazole gradient for FPLC-based His-tag purification; buffer component for enzyme assays requiring imidazole; pH calibration standard (imidazole pKa 6.95). |
| Principle / Technology | Imidazole ring nitrogen coordinates with immobilized metal ions (Ni²⁺, Co²⁺, Zn²⁺, Cu²⁺) on IMAC resin, competing with the polyhistidine tag for metal binding sites; at low concentrations (5-20 mM) imidazole reduces non-specific binding; at high concentrations (150-500 mM) imidazole elutes His-tagged proteins |
| Detection Method | Dilute 2 M stock with appropriate buffer (typically PBS or Tris-HCl with 300 mM NaCl) to desired imidazole concentration; typical wash buffer: 20-50 mM imidazole; typical elution buffer: 150-500 mM imidazole |
| Components / Formulation | Imidazole (CAS 288-32-4), HCl for pH adjustment to 7.0, ultrapure water; 0.22 µm filtered |
| Storage Conditions | 15-25 °C; protect from light (imidazole solutions may yellow slightly with prolonged light exposure); keep tightly closed |
| Shelf Life | 24 months from manufacture date; use within 12 months after opening |
| Package Specifications | 50 mL, 100 mL, 500 mL HDPE bottles |
| Product Form | Clear, colorless to very pale yellow liquid, ready-to-use after dilution |
| Key Features | 2 M stock for convenient buffer preparation; pH precisely adjusted to 7.0; 0.22 µm filtered; low heavy metal content — no interference with IMAC; compatible with Ni-NTA, Co-NTA, and other IMAC resins; essential for His-tag protein purification workflows |
| Purity | Imidazole ≥99.5% by titration; heavy metals <5 ppm; nickel <0.1 ppm (critical for IMAC applications); DNase/RNase: not detected; endotoxin <0.05 EU/mL |
| Concentration | 2.00 ± 0.05 M imidazole, pH 7.0 ± 0.05 at 25 °C |
| Activity / Unit Definition | pH buffering capacity within ±0.05 pH units of stated value |
| Molecular Weight | Not applicable — inorganic/organic buffer salts |
| Source / Origin | Synthetic, prepared from analytical grade reagents |
| pH Range / Optimal pH | pH 7.0 ± 0.05 at 25 °C; pKa of imidazole = 6.95 at 25 °C |
| Shipping Conditions | Ambient temperature |
| Expiration Date / Stability | 24 months at 15-25 °C; 12 months after opening; discard if solution yellows significantly (oxidation products may interfere with some applications); verify pH for critical applications |
| Regulatory / Compliance | For laboratory and research use only; RUO; imidazole is an irritant at high concentrations — handle with gloves; not classified as dangerous goods; ISO 9001 |
| Compatibility | Compatible with Ni-NTA, Co-NTA, Zn-IDA, and Cu-IDA IMAC resins; compatible with standard protein buffers (PBS, Tris, HEPES) containing 150-500 mM NaCl; imidazole absorbs at 280 nm — monitor protein elution by Bradford or BCA assay rather than A280; not compatible with BCA protein assay at >50 mM imidazole — use Bradford assay instead |
| Recommended Buffer System | Use as directed; compatible with biological assay systems |
| Application Notes / Precautions | Avoid contamination; use aseptic technique for sterile products; verify pH before critical applications |
| Batch-to-Batch Consistency | pH within ±0.05 units; conductivity within ±5% of reference lot |
For research use only, not for clinical use.
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