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Cat. No.: NNMB-2 (for 2ml)

Cat. No.: NNMB-10 (for 10ml)

Description

NTA-Ni Magnetic Agarose Beads for His-Tag Protein Purification, also known as NTA-Ni agarose beads (Magarose), NTA-Ni beads, or NTA nickel beads, are prepared by covalently coupling high-quality nitrilotriacetic acid (NTA) to agarose magnetic beads. The NTA is then used to chelate divalent nickel ions (Ni2+) at four binding sites, resulting in beads that can selectively bind proteins containing His-tags in lysates from animals, plants, or microorganisms, as well as in samples such as cell lysates, serum, and ascites. These beads are employed for the purification, immunoprecipitation (IP), or co-immunoprecipitation (Co-IP) of proteins or protein complexes with His-tags. The product can tolerate 6M HCl or 8M urea.

Commonly used tags include His-tag, Flag-tag, Myc-tag, HA-tag, and GST-tag. Fusion expression with these tags facilitates the convenient detection of the target protein and proteins interacting with it. These tags are also useful for the purification of the target protein.

The His-tag consists of a peptide composed of six consecutive histidine residues (HHHHHH). By recombinant DNA technology, the nucleotide sequence of the His-tag is fused to the 5' or 3' end of the target gene, resulting in the expression of the target protein with a fused His-tag. The advantages of the His-tag include its small molecular weight (0.84 kDa), minimal interference with the target protein's function, and compatibility with various protein expression systems. His-tagged proteins can be easily detected using His-tag antibodies, and purification can be achieved using His-tag protein purification agarose beads or conventional His-tag protein purification media.

The product described is developed based on immobilized metal affinity chromatography (IMAC) technology. Nickel ions, with six coordination sites, are connected to agarose or agarose magnetic beads through chelating agents such as iminodiacetic acid (IDA), nitrilotriacetic acid (NTA), or Tris (carboxymethyl) ethylenediamine (TED). NTA-Ni beads, having 4 coordination sites, exhibit intermediate binding strength between IDA and TED, making them suitable for efficient purification and convenient applications.

NTA-Ni Magnetic Agarose Beads for His-Tag Protein Purification specifically bind to His-tagged fusion proteins and can be easily applied to the purification or immunoprecipitation of proteins or protein complexes with His-tags using magnetic separation devices like magnetic racks.

Features

• This product exhibits high stability, strong specificity, and a high binding capacity for the target protein. In comparison to most similar products, this product has a higher nickel ion coordination density, resulting in lower nickel ion dissociation. It demonstrates strong specificity in binding to proteins with His-tags. Each milliliter of the suspension contains approximately 25% agarose magnetic beads precipitation, with no less than 30 μmol/ml of NTA-Ni2+. Typically, each milliliter of beads (100% beads) can bind to >20mg of His-tagged fusion protein, with specific maximum binding capacities related to the size of the tagged protein and its molecular weight.
• This product can bind to various forms of His-tagged proteins, specifically binding to N-terminal His fusion proteins (His-Protein) and C-terminal His fusion proteins (Protein-His).
• The binding of the target protein is rapid with this product. It utilizes nickel ions chelated with NTA, with a particle size around 100 μm. The adsorption process for His proteins is typically completed within 30 minutes, and the purification or immunoprecipitation of the target protein can be accomplished within 60 minutes. Shortening the operation time effectively avoids degradation or denaturation of the target protein during prolonged operations, ensuring the activity of the target protein.
• This product allows efficient elution of His-tagged proteins. Based on the structure, biological function, and subsequent application requirements of the target protein, this product utilizes imidazole for multiple elution steps. The elution process is short, with high efficiency.

Storage

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

Precautions

• The NTA-Ni agarose magnetic beads have been tested and found to maintain efficacy after three freeze-thaw cycles, but it is still recommended to aliquot the beads to minimize freeze-thaw cycles. For frequent use, store at 4ºC.

• The product should be maintained at a pH of 6-8, and avoid high-speed centrifugation and drying. Do not leave the beads in a magnetic field for extended periods, as this may cause bead aggregation.

• During use, the concentration of buffer reagents such as Tris, HEPES, and MOPS should not exceed 100mM. The concentration of chelating agents like EDTA or EGTA should not exceed 1mM. Avoid primary amine reagents such as glycine. The concentration of reducing agents such as DTT and DTE should not exceed 1mM. The concentration of detergents like Triton, Tween, and NP-40 should not exceed 2%, and the concentration of sodium deoxycholate and CHAPS should not exceed 1%. The concentration of histidine should not exceed 20mM, calcium ions should not exceed 5mM, guanidine hydrochloride can be up to 6M, urea can be up to 8M, sodium and magnesium ions can be up to 2M, and glycerol can be up to 50%. Compatibility of other reagents not mentioned should be referenced to the above but requires experimental verification.

• Before use, resuspend the product thoroughly by gently inverting several times to ensure even mixing. Avoid vigorous vortexing.

• During purification or immunoprecipitation, it is recommended to set up positive and negative control groups.

• After collecting protein samples, purification should be completed as soon as possible, and the samples should be kept at 4ºC or on ice to slow down protein degradation or denaturation. To effectively inhibit protein degradation, appropriate amounts of protease inhibitor cocktails can be added to the protein samples.

• If using a vacuum pump or other instruments to aspirate the supernatant, be mindful of the vacuum strength to avoid drawing up the aggregated beads.

• A 0.1% concentration of non-ionic detergents (such as Triton X-100, Tween-20, or NP-40) can effectively prevent bead aggregation without affecting the bead's antibody binding efficiency.

• When preparing lysis buffers independently, be aware that high concentrations of DTT or β-mercaptoethanol may affect the binding of the product to tagged proteins.

• This product is intended for scientific research by trained professionals only and is not for clinical diagnostics or therapeutic use, not for food or drug use, and should not be stored in residential areas.

• For your safety and health, please wear a lab coat and disposable gloves during operation.

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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