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| Product Name | Amino-Functionalized Magnetic Beads |
| Catalog No. | MAGBEA-0007 |
| Description | Surface-amino magnetic polymer microspheres with primary amine (-NH2) functionality designed for flexible covalent conjugation strategies including glutaraldehyde crosslinking, NHS ester chemistry, and reductive amination. The high-density amino surface enables direct coupling of carboxyl-containing ligands via EDC/NHS activation, protein immobilization through bifunctional crosslinkers, and oligonucleotide attachment. The 1.5 µm diameter provides a large surface area for high ligand loading while maintaining rapid magnetic responsiveness. |
| Intended Use | Covalent immobilization of proteins, antibodies, oligonucleotides, and other ligands for affinity purification, immunoprecipitation, DNA-binding protein pull-down, cell separation, and biosensor surface functionalization. |
| Principle / Technology | Polystyrene-divinylbenzene superparamagnetic beads are surface-functionalized with primary amine groups through sequential polymerization and post-modification amination chemistry. The amine groups serve as nucleophilic handles for multiple conjugation routes: direct coupling to carboxyl-activated ligands via EDC chemistry, homobifunctional crosslinking with glutaraldehyde or DSS, and heterobifunctional conjugation with SMCC or sulfo-SMCC for oriented sulfhydryl coupling. The highly cross-linked polymer matrix ensures low swelling and chemical stability under diverse reaction conditions. |
| Detection Method | Ninhydrin assay and TNBS titration for amine density quantitation; DLS and Coulter counter for particle sizing; SEM for surface morphology; BCA assay for protein coupling efficiency; fluorescence microscopy for coupling uniformity; zeta potential for surface charge characterization |
| Sample Type | Proteins (BSA, IgG, enzymes), antibodies (native or periodate-oxidized for oriented coupling), peptides, carboxylated oligonucleotides, carboxyl-containing small molecules, haptens, carbohydrates with reducing ends (via reductive amination); any molecule with reactive carboxyl, aldehyde, NHS ester, or epoxide groups |
| Performance Range / Specifications | Particle diameter: 1.5 ± 0.2 µm; amine density: 0.2-0.8 mmol/g (100-400 µeq/g); protein coupling capacity: 50-150 µg IgG/mg (method-dependent); non-specific binding: <5 µg BSA/mg; magnetite content: 18-25 wt%; separation time: <10 seconds; solid content: 10-50 mg/mL |
| Sensitivity / LOD | Amino group density detectable by ninhydrin assay to <0.01 mmol/g; protein coupling efficiency >80% using glutaraldehyde method; oligonucleotide coupling density: 0.5-2 nmol/mg beads (25-mer oligo); available amine groups confirmed accessible for small and large molecule coupling |
| Specificity | Primary amine groups undergo specific reactions with aldehydes, NHS esters, epoxides, and activated carboxyls; no reactivity under neutral pH with hydroxyl, unreduced sulfhydryl, or unactivated carboxyl groups; glutaraldehyde coupling specific for primary amines; crosslinked proteins retain biological activity (typically >70% of free protein activity); minimal ligand leaching from properly coupled beads |
| Reaction Conditions / Protocol | Method A (glutaraldehyde): wash beads with PBS pH 7.4; resuspend in 5% glutaraldehyde in PBS; incubate at room temperature for 2 hours with rotation; wash thoroughly to remove excess glutaraldehyde; add protein in PBS pH 7.4; incubate 2 hours room temperature or overnight at 4°C; add sodium cyanoborohydride (optional, for Schiff base reduction); wash; block with 1 M ethanolamine pH 8.0 for 1 hour; wash; resuspend in storage buffer. Method B (EDC-mediated carboxyl coupling): activate carboxyl-containing ligand with EDC/NHS in MES pH 5.5; mix activated ligand with amino beads in PBS pH 7.4-8.0; incubate 2-4 hours; wash; quench and store. |
| Components / Formulation | Amino-functionalized superparamagnetic polymer beads, 1.5 µm, 10-50 mg/mL in deionized water or 10 mM phosphate buffer pH 7.4 with 0.02% sodium azide; bifunctional crosslinkers (glutaraldehyde, EDC) not included |
| Storage Conditions | 2-8°C in sealed container; protect from light; do not freeze (amine oxidation accelerated at subzero temperatures); vortex before use; avoid exposure to aldehydes and ketones during storage (may react with surface amines); use aseptic technique |
| Shelf Life | 24 months at 2-8°C; amine density monitored by TNBS assay at 0, 12, and 24 months (<15% loss); accelerated aging confirms shelf life; opened container: 12 months with proper handling |
| Package Specifications | 1 mL, 5 mL, 10 mL, 50 mL at 10 mg/mL and 50 mg/mL; trial packs and bulk OEM packaging available; shipped in polypropylene or glass containers |
| Product Form | Dark brown to black uniform suspension; homogeneous after 30 seconds vortexing; free of visible aggregates; characteristic amine odor if using certain buffer formulations; spherical particles by microscopy |
| Quality Control | Each lot: particle size (DLS, Coulter), amine density (TNBS or ninhydrin), protein coupling capacity (rabbit IgG via glutaraldehyde method), non-specific binding, magnetic separation time, endotoxin (<0.1 EU/mg), bioburden, pH; lot traceability maintained for all raw materials |
| Key Features | Versatile amine chemistry for multiple conjugation strategies; glutaraldehyde, EDC, and NHS ester coupling options; high amine density for dense ligand loading; 1.5 µm size balances surface area and magnetic speed; suitable for oriented antibody coupling via periodate oxidation; low non-specific protein binding after blocking |
| Purity | >99% polymer-magnetite composite; endotoxin <0.1 EU/mg; heavy metals <10 ppm; no extractable free amines above specification; residual monomers below detection limit |
| Concentration | 10-50 mg/mL (labeled concentration); verified by gravimetric analysis per lot; particle count approximately 5 × 10^10 particles per mL at 10 mg/mL |
| Activity / Unit Definition | Amine density: 0.2-0.8 mmol/g; IgG coupling: 50-150 µg/mg (glutaraldehyde); oligonucleotide coupling: 0.5-2 nmol/mg; coupling efficiency >80% |
| Molecular Weight | Not applicable for polymer-magnetic composite microspheres; cross-linked polymer matrix prevents molecular weight determination |
| Source / Origin | Fully synthetic; polystyrene-divinylbenzene matrix by emulsion polymerization; magnetite encapsulation during polymerization; amine functionalization by post-polymerization chemical modification (aminolysis/reduction); all chemicals are synthetic grade; no animal-derived reagents |
| pH Range / Optimal pH | Stable: pH 3.0-12.0; amine groups protonated below pH 9-10 (pKa ~9-10 for aliphatic amines); optimal glutaraldehyde coupling: pH 7.0-8.5; optimal NHS ester coupling: pH 7.0-9.0; optimal reductive amination: pH 6.0-8.0; bead integrity lost below pH 2 or above pH 13 |
| Shipping Conditions | Ambient temperature; cold packs for summer months; protect from freezing; non-hazardous for transport; no special shipping requirements; stable during transit up to 30 days |
| Expiration Date / Stability | 24 months at 2-8°C; real-time stability at 0, 6, 12, 18, 24 months: amine density, coupling capacity, and magnetic properties all within specification; opened bottle: 12 months when handled aseptically and stored at 2-8°C; sodium azide preservative maintains antimicrobial efficacy through expiry |
| Regulatory / Compliance | For research use and further manufacturing; not intended for direct diagnostic or therapeutic use; manufactured under ISO 9001; animal-origin-free; REACH and RoHS compliant; DMF available for commercial supply agreements |
| Compatibility | Glutaraldehyde coupling: compatible with PBS, HEPES, carbonate buffers; EDC/NHS-mediated carboxyl coupling: compatible with MES activation buffer and PBS coupling buffer; NHS ester coupling: PBS, borate, carbonate buffers; avoid amine-containing buffers (Tris, glycine) during NHS ester coupling; coupled beads stable in wide range of buffers with 0.01-0.1% Tween-20 |
| Recommended Buffer System | Storage: deionized water or 10 mM sodium phosphate pH 7.4 with sodium azide; glutaraldehyde coupling: PBS pH 7.0-8.0; EDC-mediated coupling: MES pH 5.0-6.0 (activation), PBS pH 7.4-8.5 (coupling); storage after coupling: PBS pH 7.4 with 0.02% NaN3 and 0.1% BSA |
| Application Notes / Precautions | Fresh glutaraldehyde (EM grade, <1 month old) is critical for efficient coupling; remove excess crosslinker thoroughly before protein addition; sodium cyanoborohydride reduction enhances bond stability but may not be necessary for all applications; for pH-sensitive proteins, use NHS ester conjugation with DSS or BS3 crosslinkers; amino bead reactivity decreases slowly over time due to amine oxidation—use within opened shelf life; avoid carbonyl-containing buffers (ketones, aldehydes) during storage |
| Batch-to-Batch Consistency | Amine density CV <15% between batches; protein coupling capacity CV <12%; particle size CV <10%; magnetic separation time within ±3 seconds across all lots; endotoxin <0.1 EU/mg every batch; process capability (Cpk) >1.33 for critical quality attributes |
For research use only, not for clinical use.
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