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| Product Name | Carboxyl-Functionalized Magnetic Beads (1 µm) |
| Catalog No. | MAGBEA-0005 |
| Description | One-micrometer carboxyl-functionalized superparamagnetic polymer beads designed for covalent coupling of proteins, antibodies, peptides, and other biomolecules via carbodiimide-mediated amide bond formation. The high-density carboxyl surface (-COOH) provides abundant reactive sites for EDC/sulfo-NHS activation chemistry, enabling efficient and oriented biomolecule immobilization. The 1 µm particle size offers an excellent surface-area-to-volume ratio for binding capacity while maintaining sufficient magnetic content for rapid separation in chemiluminescent immunoassay and affinity purification applications. |
| Intended Use | Covalent immobilization of proteins, antibodies, antigens, enzymes, and amine-containing ligands for chemiluminescence immunoassays, ELISA development, immunoprecipitation, protein pull-down, cell separation, and custom affinity matrix preparation. |
| Principle / Technology | Uniform 1 µm superparamagnetic polystyrene-divinylbenzene beads with high-density surface carboxyl groups. Biomolecule coupling proceeds via standard EDC/NHS chemistry: EDC activates surface carboxyls to reactive O-acylisourea intermediates, which are stabilized by NHS to form semi-stable NHS-esters. Primary amines on proteins or ligands react with activated esters to form stable amide bonds. The highly cross-linked polymer matrix provides low non-specific binding and excellent chemical stability. |
| Detection Method | Dynamic light scattering and Coulter counter for particle sizing; SEM/TEM for morphology; titration for surface carboxyl density; BCA or Bradford assay for protein coupling efficiency; zeta potential for surface charge characterization; chemiluminescence intensity for functional performance in immunoassay format |
| Sample Type | Proteins, antibodies (IgG, IgM, IgA), streptavidin, Protein A/G, antigens, enzymes (HRP, ALP), peptides, and any biomolecule with primary amine groups available for covalent coupling; activation buffer: MES pH 5.0-6.0; coupling buffer: PBS or borate buffer pH 7.0-9.0 |
| Performance Range / Specifications | Particle diameter: 1.0 ± 0.1 µm (CV <10%); surface carboxyl density: 0.5-1.5 mmol/g (200-600 µeq/g); binding capacity: 80-200 µg IgG per mg beads (orientation- and buffer-dependent); non-specific binding: <5 µg BSA per mg beads; magnetite content: 15-25% by weight; separation time: <10 seconds; solid content: 10-50 mg/mL |
| Sensitivity / LOD | Detectable antibody coupling at levels as low as 0.1 µg antibody per mg beads; surface saturation achievable at 100-200 µg IgG per mg; consistent coupling efficiency >85% across lot range for recommended protocols |
| Specificity | Highly specific covalent coupling through primary amines; minimal physical adsorption (non-specific binding <1% of total binding); no reactivity with hydroxyl, sulfhydryl, or carboxyl groups without activation; coupling blocked by primary amine-containing buffer components (Tris, glycine) unless removed before activation; low batch-to-batch variation in ligand density |
| Reaction Conditions / Protocol | Wash beads twice with activation buffer (50 mM MES pH 5.5); resuspend in activation buffer; add EDC to 5-50 mM final (typically 10 mM) and sulfo-NHS to 5-50 mM (typically 10 mM); incubate at room temperature for 15-30 minutes with end-over-end rotation; wash beads twice with cold coupling buffer (PBS pH 7.4); resuspend in coupling buffer; add protein/ligand (10-200 µg per mg beads); incubate at room temperature for 2 hours or 4°C overnight with rotation; wash beads; quench residual active groups with ethanolamine or Tris (50 mM, pH 8.0) for 30 minutes; wash and resuspend in storage buffer; total coupled beads ready in 3-5 hours |
| Components / Formulation | Carboxyl-functionalized superparamagnetic polymer beads, 1 µm, suspended at 10-50 mg/mL in deionized water or 10 mM sodium phosphate pH 7.4 with 0.02% sodium azide; EDC, NHS/sulfo-NHS, activation and coupling buffers not included |
| Storage Conditions | 2-8°C in tightly sealed container; do not freeze; protect from light; store upright to minimize cap contamination; vortex before each use; use aseptic technique for long-term storage after opening |
| Shelf Life | 24 months at 2-8°C; carboxyl group stability verified by titration at 0, 12, and 24 months; accelerated aging at 37°C shows <10% loss of active carboxyls after 30 days |
| Package Specifications | 1 mL, 5 mL, 10 mL, 50 mL suspensions at 10 mg/mL and 50 mg/mL; trial pack (0.5 mL) available; bulk and OEM packaging options; supplied in polypropylene bottles with secure caps |
| Product Form | Dark brown to black homogeneous suspension; beads resuspend completely with vortexing; uniform 1 µm spheres verified by SEM; smooth surface with high-density carboxyl functionality |
| Quality Control | QC parameters: particle size by Coulter Multisizer and DLS, carboxyl density by conductometric titration, magnetic content by TGA, coupling efficiency with rabbit IgG standard (≥80 µg/mg), non-specific binding, endotoxin (<0.1 EU/mg), sterility (bioburden), zeta potential; each lot tested and certified |
| Key Features | High-density carboxyl surface for efficient coupling; uniform 1 µm size for consistent performance; low non-specific binding polymer matrix; rapid magnetic separation with high magnetite loading; EDC/NHS chemistry compatible; stable amide bond for minimal ligand leakage; suitable for chemiluminescence platform development |
| Purity | >99% polymer-magnetite composite; endotoxin <0.1 EU/mg; heavy metals within pharmaceutical limits; extractables <0.1% by weight; no detectable leachable iron under storage conditions |
| Concentration | 10-50 mg/mL as labeled; solids content verified gravimetrically per lot; particle count approximately 10^10-10^11 particles per mL at 10 mg/mL |
| Activity / Unit Definition | Protein coupling capacity: 80-200 µg IgG/mg beads; carboxyl density: 0.5-1.5 mmol/g; coupling efficiency >85% for standard EDC/NHS protocol; activity of coupled enzyme (HRP) >70% retained after immobilization |
| Molecular Weight | Not applicable for cross-linked polymer-magnetic composite particles; each bead estimated molecular weight >10^10 g/mol |
| Source / Origin | Wholly synthetic; polymer matrix produced by emulsion polymerization of styrene and divinylbenzene; magnetite nanocrystals synthesized by co-precipitation and encapsulated during polymerization; carboxyl groups introduced by copolymerization with acrylic acid or methacrylic acid monomers; all chemicals are synthetic grade |
| pH Range / Optimal pH | Stable bead suspension: pH 3.0-12.0; optimal EDC activation: pH 4.5-6.0 (MES buffer); optimal amine coupling: pH 7.0-9.0; carboxyl groups ionized above pH 5.0 (pKa ~4.5-5.0); polymer degradation onset above pH 13 |
| Shipping Conditions | Ambient temperature shipping acceptable; cold packs for summer transit in hot climates; do not freeze; non-hazardous material; standard courier service; stable during transit for up to 4 weeks |
| Expiration Date / Stability | 24 months at 2-8°C confirmed by real-time testing; carboxyl density stable (≥90% of initial) throughout shelf life; polymer matrix maintains structural integrity; magnetic responsiveness unchanged; preservative efficacy verified at multiple time points |
| Regulatory / Compliance | For research use and IVD reagent manufacturing; not for direct diagnostic use as supplied; manufactured in ISO 9001:2015 and ISO 13485 certified facility; no materials of animal origin; TSE/BSE free certification; REACH and RoHS compliant |
| Compatibility | Compatible with EDC, DCC, and other carbodiimide coupling chemistries; compatible with MES, PBS, borate, and carbonate coupling buffers; avoid Tris, glycine, and other primary amine buffers during activation step; coupled beads stable in wide range of immunoassay buffers including Tris, PBS, and HEPES |
| Recommended Buffer System | Supplied in deionized water or 10 mM sodium phosphate pH 7.4 with sodium azide; activation buffer recommended: 50 mM MES pH 5.0-6.0; coupling buffer recommended: 50-100 mM PBS or borate pH 7.4-8.5; storage buffer for coupled beads: PBS pH 7.4 with 0.02% sodium azide and 0.1% BSA (optional) |
| Application Notes / Precautions | Activate beads immediately before protein coupling (NHS-ester half-life approximately 1-2 hours in aqueous buffer); use freshly prepared EDC and NHS solutions; desalt protein into amine-free coupling buffer before bead addition; avoid over-coupling which may cause protein denaturation and reduced activity; optimize protein-to-bead ratio for each application; quench residual active groups to prevent non-specific binding in subsequent steps; coupled beads may be stored for 6-12 months at 2-8°C with appropriate preservatives |
| Batch-to-Batch Consistency | Particle size CV <10% batch-to-batch; carboxyl density CV <12%; IgG coupling capacity CV <10%; magnetic separation time within ±2 seconds across lots; endotoxin <0.1 EU/mg for all lots; comprehensive lot-to-lot quality data available |
For research use only, not for clinical use.
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