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| Product Name | Surface Plasmon Resonance (SPR) Sensor Chip, Carboxymethyl Dextran (CM5), Research Grade, 10 Chips |
| Catalog No. | PTR-HMM-0094 |
| Description | Carboxymethyl dextran (CM5) sensor chips for surface plasmon resonance (SPR) biosensor instruments (Biacore and compatible systems), designed for covalent immobilization of proteins, peptides, nucleic acids, carbohydrates, and small molecules via amine coupling, thiol coupling, or aldehyde coupling chemistry. The CM5 chip features a carboxymethylated dextran hydrogel matrix (~100 nm thickness) covalently attached to a gold-coated glass slide. The dextran matrix provides a flexible, hydrophilic three-dimensional environment for ligand immobilization, enhancing binding capacity and reducing non-specific binding compared to planar surfaces. The carboxymethyl groups are activated using standard EDC/NHS chemistry to form reactive NHS esters, which then react with primary amines on the ligand to form stable amide bonds. Each chip has 4 flow cells, allowing for reference subtraction and multi-ligand analysis. Research-grade chips are packaged in individual blister packs under inert atmosphere for long-term stability. |
| Intended Use | SPR biosensor analysis for: real-time, label-free measurement of biomolecular interaction kinetics (ka, kd, KD) and affinity (steady-state); protein-protein, protein-DNA, protein-small molecule, antibody-antigen, receptor-ligand interactions; epitope mapping and binning of antibodies; concentration analysis using calibration-free concentration analysis (CFCA) or standard curve methods; fragment-based drug screening and hit validation; and quality control of biopharmaceuticals (binding activity assays). |
| Principle / Technology | The CM5 sensor chip consists of a glass slide coated with a thin gold film (~50 nm). A self-assembled monolayer of alkanethiols anchors the carboxymethylated dextran hydrogel to the gold surface. The dextran matrix extends approximately 100 nm from the gold surface into the flow cell volume. Carboxyl groups in the dextran are activated by EDC/NHS injection to form amine-reactive NHS esters. Upon injection of the ligand (protein), primary amines react with NHS esters to form covalent amide bonds, immobilizing the ligand within the hydrogel. Ethanolamine injection deactivates remaining NHS esters. When analyte flows over the immobilized ligand, binding events cause a change in refractive index at the sensor surface, detected as a shift in the SPR angle (reported in resonance units [RU]; 1 RU approximately 1 pg/mm^2 bound protein). |
| Detection Method | 1) Dock chip in Biacore or compatible SPR instrument; 2) Prime system with running buffer; 3) Activate flow cell(s) with EDC/NHS (7 min injection); 4) Inject ligand in immobilization buffer (sodium acetate pH 4.0-5.5) at optimized concentration (typically 10-50 ug/mL) to desired immobilization level; 5) Deactivate with ethanolamine (7 min injection); 6) Condition surface with repeated analyte injections until baseline stable; 7) Perform kinetic/affinity analysis: inject analyte concentration series, allow dissociation, regenerate surface between cycles; 8) Fit data to appropriate binding model. |
| Sample Type | Ligand: purified protein (>95% purity, 10-100 ug/mL in immobilization buffer), peptide, nucleic acid, or small molecule with reactive amine. Analyte: protein, peptide, small molecule, or other binding partner in running buffer. |
| Performance Range / Specifications | Dextran matrix thickness: approximately 100 nm; carboxyl group density: approximately 1 mmol/g dextran (approximately 10^12 carboxyl groups/cm^2); protein immobilization capacity: 5-50 ng protein/mm^2 (5,000-50,000 RU) depending on protein MW; refractive index resolution: <0.1 RU (typical Biacore instruments); baseline drift: <0.3 RU/min after equilibration; chip storage: 24 months at -20 C under inert gas. |
| Sensitivity / LOD | SPR detection: <0.1 RU (approximately 0.1 pg/mm^2 protein); kinetic range: ka 10^3-10^7 M^-1 s^-1, kd 10^-5-0.5 s^-1, KD 10^-4-10^-12 M (measurable); molecular weight detection: >100 Da (small molecules) to >10^6 Da (large complexes). |
| Specificity | EDC/NHS amine coupling is specific to primary amines (N-terminus, lysine epsilon-amino group); ethanolamine deactivation prevents further non-specific binding to the dextran matrix; reference flow cell subtraction removes bulk refractive index changes, non-specific binding, and buffer mismatches; specificity of interaction determined by experimental design (competitive binding, mutant analysis, orthogonal methods). |
| Reaction Conditions / Protocol | Immobilization at 25 C (or user-specified temperature); EDC/NHS; activation: 7 min; ligand injection: typically 3-7 min at 10 uL/min; ethanolamine: 7 min; analyte injection: association typically 60-600 s, dissociation 60-3,600 s; regeneration: 30-60 s injection of appropriate solution (glycine pH 2.0-3.0, NaOH, SDS, or high salt); total cycle time: 5-20 min per concentration. |
| Components / Formulation | CM5 Sensor Chips (10 chips, each individually blister-packed under argon), Chip Storage Container, Certificate of Analysis (lot-specific carboxyl density and performance test). |
| Storage Conditions | Store unopened blister packs at -20 C; after opening, store blister pack at 2-8 C in sealed container with desiccant, use within 2 weeks; do not freeze after opening (condensation damages chip). |
| Shelf Life | 24 months from date of manufacture at -20 C unopened; 2 weeks at 2-8 C after opening blister pack. |
| Package Specifications | 10 individually packaged CM5 sensor chips; each chip has 4 flow cells. |
| Product Form | Gold-coated glass slide (approximately 12 x 12 x 0.3 mm) with carboxymethylated dextran hydrogel on one surface; mounted in plastic cassette. |
| Quality Control | Each lot tested: carboxyl group density (0.8-1.2 mmol/g dextran); dextran thickness (90-110 nm by ellipsometry); non-specific binding of BSA (1 mg/mL): <10 RU; amine coupling capacity (anti-beta-galactosidase antibody, 30 ug/mL, pH 5.0): 5,000-15,000 RU; baseline stability: <0.3 RU/min drift; surface uniformity: <5% coefficient of variation across surface. |
| Key Features | CM5 carboxymethyl dextran matrix; 100 nm hydrogel for high binding capacity; amine, thiol, and aldehyde coupling compatible; 4 flow cells per chip; individually blister-packed under argon; research-grade quality; compatible with all Biacore and most commercial SPR instruments. |
| Purity | Gold film purity: >99.9% Au; dextran: pharmaceutical grade; carboxymethylation level controlled; no detectable extractables by LC-MS. |
| Concentration | Carboxyl group density: 0.8-1.2 mmol/g dextran; protein immobilization capacity 5-50 ng/mm^2 (approximately 5,000-50,000 RU); 1 RU approximately 1 pg/mm^2. |
| Activity / Unit Definition | EDC/NHS activation efficiency: >80% of carboxyl groups activated (as measured by ligand coupling yield); anti-beta-galactosidase activity retained after immobilization: >70% of solution activity. |
| Molecular Weight | Dextran: molecular weight 500,000-1,000,000 Da before crosslinking; carboxymethyl substitution: 1 per 2-3 glucose units. |
| Source / Origin | Glass substrate: borosilicate float glass; gold: sputter-coated, >99.9% purity; dextran: from Leuconostoc mesenteroides fermentation, carboxymethylated and crosslinked with epichlorohydrin; manufactured in ISO Class 5 cleanroom. |
| pH Range / Optimal pH | Dextran matrix stable at pH 2.0-12.0; amine coupling optimal at pH 4.0-5.5 (below protein pI for electrostatic pre-concentration); analyte binding optimal at pH 7.0-8.0; regeneration pH 1.5-12.0 (short pulses). |
| Shipping Conditions | Ambient temperature in inert atmosphere blister pack; cold pack optional. |
| Expiration Date / Stability | 24 months at -20 C unopened; after opening blister pack: 2 weeks at 2-8 C desiccated; once docked in instrument: typically 1-4 weeks of continuous use (properly stored in running buffer at 2-8 C when not in use). |
| Regulatory / Compliance | For research use only; not for diagnostic or therapeutic use. |
| Compatibility | Compatible with Biacore X, X100, 3000, T100, T200, S200, 8K, and 8K+ instruments. Compatible with GE/Cytiva Biacore reagents (Amine Coupling Kit, HBS-EP+ buffer). Compatible with running buffers: HBS-EP+ (10 mM HEPES pH 7.4, 150 mM NaCl, 3 mM EDTA, 0.05% Surfactant P20), PBS, TBS, and custom buffers. DMSO up to 10% tolerated for small molecule solubility. Compatible with coupling chemistries: amine (EDC/NHS), thiol (PDEA), aldehyde (hydrazine surface modification), and streptavidin capture of biotinylated ligands (via pre-immobilized streptavidin on CM5). Not compatible with NaOH concentration >100 mM (long-term dextran hydrolysis) or 100% organic solvents (dextran matrix collapse). |
| Recommended Buffer System | Running buffer: HBS-EP+ pH 7.4 (10 mM HEPES, 150 mM NaCl, 3 mM EDTA, 0.05% Surfactant P20); Immobilization buffer: 10 mM sodium acetate pH 4.0, 4.5, 5.0, or 5.5 (choose pH below protein pI for electrostatic pre-concentration). |
| Application Notes / Precautions | Store chips at -20 C until use. Allow blister pack to reach RT before opening (approximately 30 min) to prevent condensation on the cold chip surface. Handle chip by edges only; do not touch gold or dextran surface. Dock chip according to instrument manufacturer protocol; normalize and prime before use. For amine coupling, perform a pre-concentration test (scouting) to determine optimal immobilization pH: inject ligand at 10-50 ug/mL in sodium acetate at pH 4.0, 4.5, 5.0, and 5.5 without activation; choose pH that gives +2x to +5x baseline shift (adequate electrostatic attraction for efficient coupling). Typical immobilization target: 100-500 RU for kinetics, 500-2,000 RU for concentration analysis, 5,000-15,000 RU for LMW (low molecular weight) analyte detection. The reference flow cell should undergo mock activation/deactivation without ligand. Regenerate surface between analyte injections using short pulses of appropriate solution; test regeneration conditions to ensure complete removal of analyte without loss of ligand activity. After use, store chip docked in instrument at 2-8 C with running buffer in flow cells. |
| Batch-to-Batch Consistency | Carboxyl density 0.8-1.2 mmol/g for every lot; dextran thickness 90-110 nm; baseline drift <0.3 RU/min; non-specific BSA binding <10 RU; antibody immobilization 5,000-15,000 RU. |
For research use only, not for clinical use.
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