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Cat. No.: INMB-2 (for 2ml)

Cat. No.: INMB-10 (for 10ml)

Description

IDA-Ni Magnetic Agarose Beads for His-Tag Protein Purification, also known as IDA-Ni His-tag protein purification magnetic beads, IDA-Ni magnetic agarose beads (Magarose), IDA-Ni beads, or IDA nickel beads, are composed of high-quality iminodiacetic acid (IDA) covalently coupled to agarose beads. These beads are further prepared by chelating divalent nickel ions (Ni2+) at the three binding sites of IDA. They can selectively bind to proteins containing His-tags present in samples such as cell lysates, serum, ascites, and other biological fluids, enabling the purification, immunoprecipitation (IP), or co-immunoprecipitation (Co-IP) of His-tagged proteins or protein complexes. The product is resistant to 6M guanidine hydrochloride or 8M urea.

His-tag, Flag-tag, Myc-tag, HA-tag, and GST-tag are among the most common fusion tags. These tags are fused to the target protein via recombinant gene technology, facilitating the convenient detection of the target protein and its interacting partners. They also enable easy purification of the target protein.

A His-tag consists of a peptide sequence of six consecutive histidine residues (HHHHHH). By genetically fusing the nucleotide sequence encoding the His-tag to the 5' or 3' end of the target gene, the resulting fusion protein carries the His-tag. His-tag offers several advantages: its small molecular weight (0.84 kDa) usually minimizes interference with the target protein's function and downstream applications; it typically does not form dimers and does not impact the protein's functionality; detection and localization of the target protein can be facilitated using anti-His tag antibodies; and purification can be achieved using Ni-affinity chromatography or other affinity methods.

His-tagged proteins are often purified using Ni-columns, but for applications involving the purification or immunoprecipitation of small amounts of His-tagged proteins or protein complexes, the use of His-tag protein purification magnetic agarose beads provides a simpler and more convenient alternative.

This product is developed based on the principle of immobilized metal affinity chromatography (IMAC). A nickel ion has six coordination sites, and the bridge between it and agarose beads or magnetic agarose beads is usually a chelating agent such as IDA (iminodiacetic acid), NTA (nitrilotriacetic acid), or TED (tris(carboxymethyl) ethylenediamine). IDA, NTA, and TED have 3, 4, and 5 coordination sites for binding nickel ions, respectively. This results in 3, 2, and 1 available binding sites for histidine tags. Therefore, IDA exhibits the strongest binding affinity and highest capacity for His-tagged proteins, but with lower specificity, and can be efficiently eluted using low concentrations of imidazole, while TED behaves in the opposite manner. NTA falls between the two in terms of interaction strength with His-tagged proteins.

IDA-Ni Magnetic Agarose Beads for His-Tag Protein Purification selectively bind to fusion proteins with His-tags and can be conveniently used for the purification or immunoprecipitation of His-tagged fusion proteins or their complexes using magnetic separation devices like magnetic racks.

Each milliliter of suspension of this product contains approximately 0.25 ml of agarose magnetic bead precipitate. The volume indicated for this product refers to the total volume of the suspension. Each milliliter of suspension can be used to purify 5-10 mg of His-tagged protein with a molecular weight of approximately 60 kDa. The molecular weight of the His-tagged protein will affect the amount in milligrams of the target protein that can be purified using this product.

Features

• This product exhibits high stability, strong specificity, and a high binding capacity for target proteins. Compared to most similar products, this product has a high density of nickel ion ligands, resulting in low levels of nickel ion leakage and providing strong specificity for binding to His-tagged proteins. Each milliliter of the suspension contains approximately 25% agarose magnetic beads, with no less than 30-50 μmol/ml of IDA-Ni2+. Typically, each milliliter of magnetic beads (100% beads) can bind to 20-40 mg of His-tagged fusion protein, with the maximum binding capacity dependent on the size of the target protein and the size of the His-tag.
• This product can bind to various forms of His-tagged proteins, including N-terminal His fusion proteins (His-Protein) and C-terminal His fusion proteins (Protein-His).
• The binding process of the target protein using this product is rapid. The product employs nickel ions chelated with IDA and has a particle size of around 100 μm. Typically, the adsorption of His-tagged proteins can be completed within 30 minutes, and the purification or immunoprecipitation of the target protein can be achieved within 60 minutes. Shortening the operation time effectively prevents protein degradation or denaturation during prolonged procedures, ensuring the activity of the target protein.
• This product enables efficient elution of His-tagged proteins. Depending on the structure of the target protein, its biological function, and the requirements of subsequent applications, multiple elutions are performed using imidazole. The elution process is short and efficient.
• This product is convenient to use. Stored in a special protective solution without glycerol, it allows for rapid and efficient separation through magnetic adsorption, eliminating the need for centrifugation steps.

Storage

Store at -20ºC, valid for two years. Store at 4ºC, valid for one year.

Related: 

• TED-Ni Magnetic Agarose Beads for His-Tag Protein Purification
• IDA-Ni Magnetic Agarose Beads for His-Tag Protein Purification
• NTA-Ni Magnetic Agarose Beads for His-Tag Protein Purification

Only for research and not intended for treatment of humans or animals

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