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  • In general, the term immobilization refers to the act of the limiting movement or making incapable of movement. Immobilization is a universal phenomenon existing in nature. In the fields of biological and biomedical, many molecules such as biopolymers and small molecules are artificially immobilized on solid support as important research tools. The solid support should exhibit good chemical and physical stability and contain functional groups that can be bound to the active molecule. Common solid supports are magnetic particles, agarose beads, nanoparticles, films, microplates, array surfaces, and strips. The main methods for immobilization are adsorption, covalent binding, entrapment, encapsulation, and crosslinking, and the appropriate method should be selected according to the research purpose. The formation of covalent bonds between the ligand and the support usually provides the stronger linkages compared with other immobilization methods. The types of functionalities used for ligand attachment are easily reactive components such as amines, sulfhydryls, aldehydes, and carboxylic acids.

    Product Range

    Amerigo Scientific offers SepSphere™ kits for the immobilization of small molecules and large biomolecules (including proteins, antibodies, peptides, and oligonucleotides) on agarose beads. All processes of immobilization, washing of unreacted starting materials, and column packing are performed in a column. The required consumables and reagents are available in the SepSphere™ kit. The washing is performed using a simple manual push syringe without the need for gas/air flow or vacuum.

    Product Ligand Ligand Reactive Groups Agarose Reactive Groups Linkage Stability of Linkage
    SepSphere™ Antibody/Protein/Enzyme Immobilization Kit via Glyoxyl Antibody/Protein/Enzyme -NH2 -C(=O)H (glyoxyl) Secondary amine Stable
    SepSphere™ Antibody/Protein/Enzyme Immobilization Kit via Glutaraldehyde Antibody/Protein/Enzyme -NH2 Glutaraldehyde Imine Stable at pH 5-9
    SepSphere™ Peptide Immobilization Kit via Maleimide Peptide -SH (Cys thiol) Maleimide Thiol ether Stable
    SepSphere™ Peptide Immobilization Kit via Pyridyl Disulfide Peptide -SH (Cys thiol) -S-S-Pyridine Disulfide bond Stable*
    SepSphere™ Peptide Immobilization Kit via Carboxylic Acid Peptide -NH2 -COOH Amide bond Stable
    SepSphere™ Oligo Immobilization Kit via Carboxylic Acid Oligo -NH2 -COOH Amide bond Stable
    SepSphere™ Oligo Immobilization Kit via Epoxide Oligo -NH2 -CH(O)CH2 (epoxide) β-amino alcohol Stable
    SepSphere™ Oligo Immobilization Kit for Phosphate DNA Oligo -OPO3 (phosphate) -COOH Amide bond Stable
    SepSphere™ Small Molecule Acid Immobilization Kit Small molecule -COOH -NH2 Amide bond Stable
    SepSphere™ Small Molecule Amine Immobilization Kit via Glyoxyl Small molecule -NH2 -C(=O)H (glyoxyl) Secondary amine Stable
    SepSphere™ Small Molecule Amine Immobilization Kit via Epoxide Small molecule -NH2 -CH(O)CH2 (epoxide) β-amino alcohol Stable
    SepSphere™ Small Molecule Alcohol Immobilization Kit Small molecule -OH (aliphatic alcohol) -NH2 Thiol ether Stable
    SepSphere™ Small Molecule Amine Immobilization Kit via Carboxylic Acid Small molecule -NH2 -COOH Amide bond Stable
    SepSphere™ Small Molecule Ketone Immobilization Kit Small molecule -C=O- (Ketone or aldehyde) -ONH2 (aminooxy) Oxime bond Stable*

    * Releasable under reducing conditions

    Features of SepSphere™ Immobilization Kits

    • All reagents, buffers, and consumables for immobilization included in the kits
    • Agarose beads with high loading capacity and stability
    • Pre-made buffers for easy direct use
    • Immobilization, washing, and packing all performed within a column
    • Minimum requirement for immobilization set-up

    Design of Columns in SepSphere™ Immobilization Kits

    The column in the SepSphere™ kit is the core component of molecular immobilization. The column consists of five parts.

    Design of Columns in SepSphere™ Immobilization Kits

    1. Male luer lock cap for facilitating fluid mixing
    2. Column top and luer attachment. The luer attachment is used as a tight seal and attachment of the syringe for washing. The column top can be removed to allow buffer addition and agitation.
    3. Column body containing a polypropylene frit at the bottom
    4. Additional frit for column packing after immobilization
    5. Bottom female luer lock plug for facilitating fluid mixing and storage
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