Elution Buffer For Ni-NTA/Ni-IDA,250 mM Imidazole pH 7.9~8.1

Description

Biofargo Elution Buffer for Ni-NTA / Ni-IDA (250 mM Imidazole, pH 7.9-8.1) is a ready-to-use elution buffer optimized for efficient and selective recovery of His-tagged recombinant proteins in immobilized metal affinity chromatography (IMAC) workflows. Formulated with 250 mM imidazole at near-neutral pH, this elution buffer competitively displaces His-tagged proteins bound to Ni-NTA and Ni-IDA resins, resulting in high yield and high purity eluted fractions.

Designed for compatibility with gravity flow columns, syringes, spin columns, and automated FPLC systems, this buffer provides a consistent, reproducible elution performance that meets the demands of modern protein purification applications in research and bioprocess development.

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Features

• High-Strength 250 mM Imidazole Formulation — Efficiently displaces His-tagged proteins from Ni-chelated resins. 

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• Neutral pH 7.9-8.1 — Maintains protein integrity and supports broad compatibility with His-tag purification workflows. 

• Compatible with Ni-NTA & Ni-IDA Resins — Works seamlessly with industry-standard IMAC materials.

• Ready-to-Use Liquid Buffer — Eliminates buffer preparation time, reduces error, and ensures consistency.

• Improves Target Recovery & Purity — Designed to elute specifically bound proteins while minimizing carry-over of contaminants. 

• Multipurpose Workflow Compatibility — Suitable for gravitational flow, spin, and FPLC chromatography systems.

Applications

This elution buffer is ideal for use in the following HIS-tag protein purification workflows:

• Competitive elution of recombinant His-tagged proteins from Ni-NTA / Ni-IDA affinity media

• High-yield recovery in native and denaturing conditions

• Gravity flow columns, spin column formats, FPLC and automated systems

• Protein expression screening and process development workflows

• Downstream analytical preparation and functional assays

Usage Instructions

Compatible with elution buffers for Ni-NTA/IDA resin purification. For better elution efficiency, gradient elution with imidazole is recommended to determine the optimal elution concentration. Generally, the imidazole concentration ranges from 20 mM to 500 mM, and 250 mM can elute most proteins. For Ni-TED resin, use with product C600501.

Components

Components include: 250 mM Imidazole, NaCl, NaH2PO4·2H2O, pH 7.9~8.1

How It Works

His-tagged proteins bind strongly to immobilized nickel ions coordinated by NTA or IDA chelators on the resin. During elution, imidazole at high concentration competes with histidine residues on the protein tag, releasing the target protein from the resin without harsh conditions.

Typical workflows include:

• Equilibration of Ni-NTA / Ni-IDA resin

• Lysate loading and wash steps

• Elution with 250 mM imidazole buffer

• Collection of purified target protein fractions