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| Product Name | Transfection Enhancer, Polyamine-Based, Post-Transfection Supplement |
| Catalog No. | ATR-CTR-0030 |
| Description | Polyamine-based transfection enhancer solution applied post-transfection to significantly boost transgene expression levels in mammalian cells transfected with lipid-based, polymer-based, or calcium phosphate reagents. The enhancer contains a proprietary blend of natural polyamines (spermine, spermidine) and a small molecule chromatin decondensation agent that synergistically increase transcriptional activity from plasmid DNA templates. Mechanism involves: (1) relaxation of chromatin structure around episomal plasmid DNA to increase promoter accessibility, (2) enhanced nuclear import of plasmid DNA through polyamine interactions with nuclear pore complex proteins, and (3) stabilization of mRNA transcripts through polyamine-ribosome interactions. Expression enhancement of 2-10 fold is typically observed when added 4-6 hours post-transfection. |
| Intended Use | Post-transfection enhancement of transient transgene expression in mammalian cells for applications requiring high-level recombinant protein production, reporter gene assays, and lentivirus/AAV production where maximum expression is desired. |
| Principle / Technology | Polyamines neutralize negative charges on DNA and chromatin, facilitating chromatin decondensation and transcription factor access; spermine interacts with nuclear pore complex Nup proteins to enhance plasmid nuclear import; polyamines stabilize mRNA secondary structure and enhance translation initiation. |
| Detection Method | Add enhancer directly to culture medium at 1:500 to 1:200 dilution 4-6 hours after transfection; incubate cells for the remaining expression period (24-72 hours); no medium change required; harvest cells or supernatant as usual. |
| Sample Type | Mammalian cell cultures post-transfection with plasmid DNA (lipid, polymer, or calcium phosphate based transfection). |
| Performance Range / Specifications | Expression enhancement: 2-10 fold increase in reporter gene activity or protein yield compared to untreated transfected cells; effective across HEK293, CHO, HeLa, COS-7, and selected primary cells. |
| Sensitivity / LOD | Enhancement detectable at 1:1000 dilution; optimal at 1:500-1:200; no cytotoxicity at recommended concentrations (cell viability >95% at 1:200 dilution, 48 hr). |
| Specificity | Enhances expression from plasmid DNA templates; does not enhance expression from integrated transgenes or endogenous genes; effect is specific to transiently transfected episomal plasmids. |
| Reaction Conditions / Protocol | Add enhancer 4-6 hours post-transfection at 1:200-1:500 dilution; incubate without medium change for remainder of expression period; peak enhancement at 24-72 hours depending on promoter and cell type. |
| Components / Formulation | Spermine, spermidine, proprietary chromatin relaxation agent, in sterile PBS, pH 7.4; 0.22 µm filtered. |
| Storage Conditions | Store at -20 °C; protect from light; avoid repeated freeze-thaw cycles; aliquot upon first thaw. |
| Shelf Life | 12 months from date of manufacture at -20 °C. |
| Package Specifications | 0.5 mL (sufficient for ~250 transfections in 6-well format), 2.5 mL (sufficient for ~1250 transfections). |
| Product Form | Clear, colorless to pale yellow liquid; sterile-filtered. |
| Quality Control | Each lot tested for expression enhancement in HEK293T cells transfected with pCMV-EGFP (2-10 fold enhancement by FACS), cytotoxicity (<5% cell death at 1:200 dilution), sterility, and endotoxin (<0.1 EU/mL). |
| Key Features | Post-transfection enhancer — no re-optimization of transfection protocol needed; 2-10 fold expression boost; natural polyamine-based, low toxicity; simple add-to-medium protocol; compatible with most transfection reagents; works across multiple cell lines and promoters. |
| Purity | Spermine ≥97%; spermidine ≥98%; chromatin agent synthetic purity ≥95%. |
| Concentration | Ready-to-use; add 2-5 µL per mL culture medium (1:500-1:200 dilution). |
| Activity / Unit Definition | Expression enhancement activity: 2-10 fold increase in EGFP fluorescence or SEAP activity vs. untreated transfected cells. |
| Molecular Weight | Spermine: 202.34 g/mol (C10H26N4); spermidine: 145.25 g/mol (C7H19N3). |
| Source / Origin | Synthetic and naturally-derived polyamines; proprietary chromatin relaxation small molecule. |
| pH Range / Optimal pH | pH 7.4 ± 0.1. |
| Shipping Conditions | Cold pack or dry ice; protect from light. |
| Expiration Date / Stability | 12 months at -20 °C; after first thaw, stable for 2 weeks at 2-8 °C. |
| Regulatory / Compliance | For research use only; not for therapeutic or diagnostic use. |
| Compatibility | Compatible with all standard mammalian cell culture media (DMEM, RPMI-1640, MEM, F-12, etc.) with or without serum. Compatible with lipid-based (Lipofectamine), polymer-based (PEI, polybrene), and calcium phosphate transfection methods. Not recommended for electroporation — add post-electroporation after cells have attached (4-6 hr). |
| Recommended Buffer System | PBS, pH 7.4. |
| Application Notes / Precautions | Optimal timing: add 4-6 hours after transfection when plasmid DNA has entered the nucleus but before peak expression. Do not add enhancer simultaneously with transfection reagent — polyamine-transfection reagent interaction may reduce transfection efficiency. For protein production, add enhancer at 4 hr and harvest at 48-72 hr (optimal). For secreted proteins, enhancer can be re-added at 24 hr for extended production. Run an enhancer-absent control to quantify fold-enhancement. Enhancer is compatible with antibiotic selection for stable cell line generation — add after transfection but before starting selection. |
| Batch-to-Batch Consistency | Polyamine content within ±10% of specification; expression enhancement in standard EGFP assay within ±20% of reference lot. |
For research use only, not for clinical use.
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