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| Product Name | Electroporation Buffer for Mammalian Cells, Optimized Conductivity |
| Catalog No. | ATR-CTR-0026 |
| Description | Specialized electroporation buffer with optimized ionic composition and conductivity for efficient nucleic acid delivery into difficult-to-transfect mammalian cells via electroporation (nucleofection). The buffer is formulated with a proprietary combination of potassium, magnesium, and phosphate ions at precisely controlled concentrations to achieve the optimal conductivity (2-5 mS/cm) that balances efficient pore formation with minimal cell damage during electric pulse application. Supplemented with ATP-regenerating components (creatine phosphate and creatine kinase) to support rapid cellular energy recovery post-electroporation, and a membrane-stabilizing polymer that promotes resealing of transient membrane pores. |
| Intended Use | Electroporation-based delivery of plasmid DNA, siRNA, mRNA, and CRISPR ribonucleoprotein (RNP) complexes into hard-to-transfect mammalian cells including primary cells, stem cells, and immune cells. |
| Principle / Technology | Precise ionic conductivity (2-5 mS/cm) creates optimal current density for reversible membrane pore formation; ATP regeneration system (creatine phosphate + CK) supports rapid cellular recovery; membrane-stabilizing polymer promotes pore resealing; potassium-based formulation mimics intracellular ionic environment during electroporation. |
| Detection Method | Resuspend 1×10^5-1×10^7 cells in 100 µL electroporation buffer; add nucleic acid (1-10 µg DNA or 10-200 pmol siRNA); transfer to cuvette; electroporate using optimized pulse program; immediately add pre-warmed culture medium; transfer cells to culture vessel. |
| Sample Type | Primary mammalian cells, stem cells (ESC, iPSC), immune cells (T cells, B cells, NK cells, dendritic cells), hard-to-transfect cell lines (Jurkat, K562, THP-1), primary neurons. |
| Performance Range / Specifications | Transfection efficiency: 30-90% depending on cell type; cell viability post-electroporation: 50-90% at 24 hours; applicable cell density: 1×10^5-2×10^7 cells/mL. |
| Sensitivity / LOD | Transgene expression detectable within 4-6 hours post-electroporation (depending on promoter strength); siRNA knockdown detectable within 24 hours. |
| Specificity | Formulation optimized for mammalian cell electroporation; not suitable for bacterial or yeast electroporation which require different ionic conditions. |
| Reaction Conditions / Protocol | Electroporation parameters (voltage, pulse length, pulse number) must be optimized for each cell type; typical square wave: 100-300 V, 1-30 msec, 1-3 pulses; exponential decay wave: 200-350 V, 500-960 µF. |
| Components / Formulation | Electroporation buffer (5× concentrate), ATP regeneration supplement (creatine phosphate, creatine kinase, ATP), membrane stabilizer (proprietary polymer), nuclease-free water, protocol. |
| Storage Conditions | Store at 2-8 °C; ATP supplement at -20 °C; protect from light. |
| Shelf Life | 12 months from date of manufacture. |
| Package Specifications | 25 reactions (2.5 mL 5× buffer), 100 reactions (10 mL), 500 reactions (50 mL). |
| Product Form | 5× concentrated buffer: clear, colorless liquid; ATP supplement: lyophilized powder; membrane stabilizer: liquid supplement. |
| Quality Control | Each lot tested for conductivity, osmolality, pH, sterility, endotoxin (<0.1 EU/mL), and transfection efficiency in Jurkat T cells (pmaxGFP plasmid, >50% GFP+). |
| Key Features | Optimized conductivity for efficient electroporation; ATP regeneration system for improved viability; membrane stabilizer for pore resealing; validated for primary cells and stem cells; compatible with DNA, RNA, and RNP delivery; RNase/DNase-free. |
| Purity | Buffer components analytical grade; ATP supplement >99% purity; endotoxin <0.05 EU/mL. |
| Concentration | Dilute 5× buffer to 1× with nuclease-free water before use. |
| Activity / Unit Definition | Conductivity: 2-5 mS/cm (1×); membrane pore resealing >80% within 5 minutes post-electroporation as measured by propidium iodide exclusion recovery. |
| Molecular Weight | Not applicable — multi-component buffer system. |
| Source / Origin | Analytical and molecular biology grade buffer salts; recombinant creatine kinase; synthetic membrane polymer. |
| pH Range / Optimal pH | pH 7.2-7.4 at 25 °C (1×). |
| Shipping Conditions | Cold pack (ATP supplement on dry ice). |
| Expiration Date / Stability | 12 months at recommended storage; after reconstitution, ATP supplement stable 1 month at -20 °C. |
| Regulatory / Compliance | For research use only; not for therapeutic or diagnostic procedures. |
| Compatibility | Compatible with Lonza Nucleofector, Bio-Rad Gene Pulser, BTX ECM, and Neon Transfection System platforms. Verify cuvette gap size (1, 2, or 4 mm) compatibility. Use with purified nucleic acid (A260/A280 1.8-2.0); endotoxin-free plasmid preparation recommended. |
| Recommended Buffer System | Potassium phosphate-based buffer with Mg2+ and ATP regeneration system, pH 7.3. |
| Application Notes / Precautions | Pre-warm culture medium to 37 °C before electroporation. After pulse, immediately add pre-warmed medium and transfer cells gently — do not pipette vigorously. Serum-free medium for the first 2-4 hours post-electroporation can improve viability for some cell types. Optimize DNA amount (1-10 µg per 1×10^6 cells) for each construct. Include no-DNA control for viability baseline. Electroporation efficiency depends on cell health — use log-phase cultures. Nucleofection programs should be optimized for each cell type. |
| Batch-to-Batch Consistency | Conductivity within ±0.5 mS/cm of reference; transfection efficiency in standard Jurkat assay within ±10% of reference lot. |
For research use only, not for clinical use.
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