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| Product Name | Carboxyl Magnetic Beads, 1 µm, for Covalent Ligand Coupling, EDC/NHS-Ready |
| Catalog No. | MAGBEA-0033 |
| Description | Carboxyl-functionalized monodisperse magnetic beads (1 µm diameter) with high surface carboxyl group density for covalent immobilization of proteins, antibodies, peptides, oligonucleotides, and small molecule ligands via EDC/NHS (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide/N-hydroxysuccinimide) chemistry. The carboxyl groups on the bead surface form stable amide bonds with primary amines on the ligand of interest when activated by water-soluble EDC and stabilized by NHS or Sulfo-NHS. The 1 µm particle size provides an optimal balance of surface area for ligand coupling, rapid magnetic separation kinetics, and suitability for microfluidic and automated liquid handling platforms. |
| Intended Use | Covalent coupling of antibodies for immunoprecipitation and immunoassay development; protein immobilization for pull-down and affinity purification; oligonucleotide coupling for DNA/RNA-binding protein capture; small molecule immobilization for drug target identification; custom affinity resin preparation. |
| Principle / Technology | Carboxyl groups on bead surface activated by EDC to form reactive O-acylisourea intermediate; NHS converts this to amine-reactive NHS ester (more stable); primary amines on ligand react with NHS ester forming stable amide bond; ligand covalently and irreversibly attached with preferred orientation (through surface-accessible lysine residues or N-terminus). |
| Detection Method | Wash beads 2× with activation buffer (MES, pH 5.0-6.0); activate with EDC (5-10 mM) + Sulfo-NHS (5-20 mM) for 15-30 min at RT with rotation; wash quickly 2× to remove excess EDC/NHS; add ligand in coupling buffer (PBS or MES, pH 7.0-8.0, no Tris/glycine/other primary amines); incubate 2 hours at RT or overnight at 2-8 °C; quench unreacted NHS esters with 1 M ethanolamine or Tris pH 8.0 for 15 min; wash and store beads. |
| Sample Type | Purified antibodies (1-10 mg/mL), recombinant proteins, synthetic peptides, amine-modified oligonucleotides, small molecule amine derivatives. |
| Performance Range / Specifications | Ligand coupling capacity: 10-50 µg IgG per mg beads (0.5-2.5 mg per mL settled beads); carboxyl group density: 0.2-0.5 mmol/g beads; coupling efficiency: 50-90% depending on ligand concentration and MW. |
| Sensitivity / LOD | Detection of immobilized ligand by BCA protein assay or direct fluorescent labeling; functional activity of coupled antibody verified by antigen binding assay. |
| Specificity | EDC/NHS chemistry specifically targets primary amines (-NH2); other nucleophiles (-SH, -OH) much less reactive with NHS ester; Tris, glycine, and other amine-containing buffers must be avoided during coupling step. |
| Reaction Conditions / Protocol | EDC/NHS activation: 15-30 min at RT pH 5-6; ligand coupling: 1-2 hours at RT or overnight at 4 °C pH 7-8; quenching: 15 min with 1 M ethanolamine pH 8. |
| Components / Formulation | Carboxyl magnetic beads (10 mg/mL suspension in water or PBS), EDC (lyophilized, 100 mg), Sulfo-NHS (lyophilized, 100 mg), MES activation buffer (0.1 M, pH 5.5), coupling buffer (PBS, pH 7.4), quenching buffer (1 M ethanolamine, pH 8.0), protocol. |
| Storage Conditions | Store carboxyl beads at 2-8 °C; do not freeze; EDC and Sulfo-NHS at -20 °C desiccated. |
| Shelf Life | 12 months from date of manufacture. |
| Package Specifications | 1 mL (10 mg) bead suspension, 5 mL (50 mg), 10 mL (100 mg); EDC + Sulfo-NHS sufficient for coupling reactions. |
| Product Form | Dark brown/black magnetic bead suspension; settles in magnetic field within 10-20 seconds. |
| Quality Control | Each lot tested for carboxyl group density (toluidine blue or conductivity titration), particle size (DLS: 1.0 ± 0.1 µm, PDI <0.1), nonspecific protein binding (<5 µg BSA/mg beads), and IgG coupling efficiency (>50 µg/mg). |
| Key Features | EDC/NHS-ready carboxyl beads; 1 µm monodisperse for high surface area; EDC and Sulfo-NHS included; stable amide bond linkage; reusability (coupled ligand beads stable for months); low nonspecific binding; compatible with automated platforms. |
| Purity | Carboxyl group density 0.2-0.5 mmol/g; iron content >80% w/w; surfactant and preservative free. |
| Concentration | 10 mg/mL bead suspension; coupling capacity 10-50 µg IgG/mg beads. |
| Activity / Unit Definition | EDC/Sulfo-NHS activation efficiency: >80% of carboxyl groups activated based on NHS ester colorimetric assay. |
| Molecular Weight | EDC: 191.70 g/mol (C8H17N3 · HCl); Sulfo-NHS: 217.13 g/mol (C4H4NNaO6S). |
| Source / Origin | Magnetic iron oxide core with carboxylated polymer shell; synthetic EDC and Sulfo-NHS. |
| pH Range / Optimal pH | Activation: pH 5.0-6.0 (MES buffer); coupling: pH 7.0-8.0; storage: pH 7.4 (PBS). |
| Shipping Conditions | Cold pack (2-8 °C); EDC/NHS on dry ice recommended. |
| Expiration Date / Stability | 12 months at 2-8 °C for beads; EDC and NHS stable at -20 °C for 12 months. |
| Regulatory / Compliance | For research use only; not for diagnostic or therapeutic use. |
| Compatibility | Beads compatible with most aqueous buffers. Avoid Tris, glycine, ammonium, and other primary amine-containing buffers during coupling step. Avoid phosphate buffers (>50 mM) during EDC activation as phosphate may react with EDC. For hydrophobic ligands, up to 10% DMSO or DMF can be added during coupling. |
| Recommended Buffer System | MES 0.1 M pH 5.5 for activation; PBS or borate buffer pH 7.4-8.5 for coupling. |
| Application Notes / Precautions | EDC and Sulfo-NHS are moisture-sensitive — aliquot upon first opening and store desiccated at -20 °C. Prepare activation reagents fresh immediately before use (EDC half-life ~2 hours in aqueous solution at pH 5-6). For optimal coupling efficiency, use 1-5 mg/mL ligand concentration. Include a coupling control where beads are activated and quenched without ligand to measure nonspecific binding. After coupling, measure protein concentration in supernatant to calculate coupling efficiency. Store coupled beads in PBS with 0.02% NaN3 at 2-8 °C. |
| Batch-to-Batch Consistency | Carboxyl group density within ±15% of specification; particle size within ±10% (DLS); IgG coupling capacity within ±20% of reference lot. |
For research use only, not for clinical use.
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