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NPT01
NeuroPorter™ Transfection Kit
Lipid formulation for nucleic acid transfections in neuronal and glial cells
Synonym(e):
NeuroPorter Transfection, Transfection Kit
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| Packungsgröße | SKU | Verfügbarkeit | Preis |
|---|---|---|---|
| 1 kit | Warenkorb auf Verfügbarkeit prüfen | CHF 775.00 |
Über diesen Artikel
NACRES:
NA.25
UNSPSC Code:
12352200
Form:
dried film
Grade:
Molecular Biology
Technique(s):
transfection: suitable
CHF 775.00
Warenkorb auf Verfügbarkeit prüfen
Technischer Dienst
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Unterstützung erhaltengrade
Molecular Biology
Quality Segment
form
dried film
usage
kit sufficient for 75-200 transfections
availability
available only in USA, Canada and EU
technique(s)
transfection: suitable
storage temp.
2-8°C
General description
Neuroporter™ Transfection Reagent is a unique formulation of a proprietary cationic lipid optimized for delivery of DNA into primary neurons, glial cells, and cultured neuronal cell lines with high efficiency and low toxicity. The Neuroporter™ Transfection Kit was designed for difficult-to-transfect primary neurons, addressing past problems such as poor cell viability, low transfection efficiency and neuro-degeneration.
Application
Suitable for transient and stable transfection of nucleic acids into primary neurons and cultured neuronal cell lines. Use approximately 15-120 μl Neuroporter™ Transfection Reagent and 6-8 μg DNA (in provided unique DNA Dilution buffer when required) per 6 cm cell culture plate. The following cells have been successfully transfected using the Neuroporter™ Transfection Kit:
- C6 glioma (human)
- Cortical neurons (rat primary)
- Dorsal Root Ganglion (DRG) cells (rat)
- NT2 neurons(human precursor cells)
- NT neurons (human differentiated cells)
- Subventricular Zone (SVZ) cells (mouse)
- White matter cells (mouse)
Biochem/physiol Actions
A stable, non-covalent complex is formed when the Neuroporter™ Transfection Reagent is mixed with DNA in the absence of serum. The complexes are stable and can be directly added to the cell culture medium, where they fuse with the cell membrane, releasing the DNA into the cytoplasm. Note: complex formation is inhibited by serum, but once stable complexes have formed, the presence of serum is without consequence.
Features and Benefits
- Optimized for primary neurons, glial cells, and cultured neural cell lines
- Very low toxicity with no neuro-degeneration or dendrite withdrawal
- Efficient DNA delivery primary neurons, glial cells, and cultured neural cell lines
- Fast and easy to use compared to other methods
- Compatible with both serum and serum-free transfection protocols
Other Notes
1 vial Neuroporter™ Transfection Reagent, dried lipid film (T2823)
1.5 mL Hydration Buffer H9036
7.5 mL DNA Diluent D1941
1.5 mL Hydration Buffer H9036
7.5 mL DNA Diluent D1941
Legal Information
NeuroPorter is a trademark of Gene Therapy Systems, Inc.
Disclaimer
Do not freeze.
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Dieser Artikel | |||
|---|---|---|---|
| form dried film | form solution | form liquid | form liquid |
| grade Molecular Biology | grade Molecular Biology | grade - | grade - |
| technique(s) transfection: suitable | technique(s) transfection: suitable | technique(s) transfection: suitable | technique(s) transfection: suitable |
| Quality Level 200 | Quality Level 200 | Quality Level 200 | Quality Level 100 |
| storage temp. 2-8°C | storage temp. −20°C | storage temp. 2-8°C | storage temp. 2-8°C |
| usage kit sufficient for 75-200 transfections | usage kit sufficient for 160 transfections (6 cm dishes), kit sufficient for 400 transfections (3.5 cm dishes), kit sufficient for 80 transfections (10 cm dishes) | usage - | usage - |
Lagerklasse
12 - Non Combustible Liquids
flash_point_f
Not applicable
flash_point_c
Not applicable
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Artikel
Transfection introduces genetic material into cells, aiding research in gene expression and cell biology.
This brief webinar provides an overview of what transfection is and the methods that are used to introduce DNA or RNA into eukaryotic cells.
Verwandter Inhalt
Browse our convenient transfection reagent selection guide to match the best reagent for your specific cell line and application needs.
Instructions
Nikhil G Thaker et al.
Journal of neuroscience methods, 185(2), 204-212 (2009-09-29)
A major challenge for the treatment of cancers, such as glioblastoma multiforme (GBM), has been resistance to radiation and cancer chemotherapeutics. Short interfering RNA (siRNA) based screening may facilitate the identification of genes and pathways essential for cancer cell survival
Simone Di Giovanni et al.
The Journal of biological chemistry, 280(3), 2084-2091 (2004-11-04)
Following spinal cord injury, there are numerous changes in gene expression that appear to contribute to either neurodegeneration or reparative processes. We utilized high density oligonucleotide microarrays to examine temporal gene profile changes after spinal cord injury in rats with
Beata Jablonska et al.
The Journal of cell biology, 179(6), 1231-1245 (2007-12-19)
We investigated the function of cyclin-dependent kinase 2 (Cdk2) in neural progenitor cells during postnatal development. Chondroitin sulfate proteoglycan (NG2)-expressing progenitor cells of the subventricular zone (SVZ) show no significant difference in density and proliferation between Cdk2(-/-) and wild-type mice
Global Trade Item Number
| SKU | GTIN |
|---|---|
| NPT01-1KT | 04061834211648 |
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What is the Department of Transportation shipping information for this product?
1 answer-
Helpful?
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Is the size of the plasmid an important consideration for transfection?
1 answer-
The size of the plasmid should be considered when selecting a transfection reagent with the best efficiency. In general, larger sized plasmids should easily transfect with readily available transfection reagents, as along as the plasmid DNA is of high purity.
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What is the difference between stable and transient transfection?
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When the DNA enters the nucleus of the cell, the plasmid is replicated by the cell machinery (transient transfection). During this time, RNA is transcribed and protein translated until the plasmid DNA is lost after a few cell divisions. This expression of the plasmid DNA, mRNA, and protein is transient (temporary).In some cases, the plasmid DNA is integrated into the host cell genome. This is usually accompanied by forced expression using a selection antibiotic and sometimes a cloning step (to be sure all cells have the same integration site). Once the DNA is stable, the cell line can be frozen and used to express protein for many years. Clones may even be screened for those expressing the highest amount of protein.
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Is optimizing the transfection protocol important?
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For many common cell lines, transfection reagent efficiency is very high and the protocols will not require any optimization. For hard-to-transfect cells or those ultimately expressing a toxic protein, the protocol should be optimized for best transfection efficiency. Taking time to optimize will give you more transfected cells with each procedure, which can mean more protein expressed and results that are reproducible.
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How can I determine the efficiency of my transfection?
1 answer-
Calculating transfection efficiency is very useful when optimizing transfection protocols. Transfection efficiency can be performed using a GFP-expressing plasmid. After transfection, cells are stained with propidium iodide and counted. The propidium iodide provides a count of the total cells in the population, and the GFP-expressing cells provide a count of the number of cells transfected. The transfection efficiency (%) can then be calculated by:(# GFP-expressing cells / total cell #) * 100
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Can antibiotics be present in the medium during transfection?
1 answer-
We recommend that no antibiotics are present during transfection. The process of transfection can make the cells somewhat more porous to allow for efficient DNA entry. During this time, antibiotics will also enter the cells more easily and the cells may show increased cell death. Wait until about 24 hours after transfection to resume the use of preventative antibiotics and/or start the use of selective antibiotics.
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What quality does the DNA need to be in order to use it for transfection?
1 answer-
The DNA needs to be good quality or it may cause the cells to lyse and/or they won't transfect efficiently. Plasmid DNA prepared with a column-based DNA purification kit is suitable for transfections. Sigma's GenElute™ Minprep, Midiprep and Maxiprep kits work well for DNA plasmid purification. After preparing the DNA, confirm the OD A260:A280 ratio is greater than 1.6 for use in plasmid transfections.
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Is low cell passage number an important consideration for transfection?
1 answer-
Yes, we recommend cells are at a low passage when being used for any application, including transfection. The reason why depends on what type of cells they are. Primary cells will undergo a finite number of divisions, and as they get closer to senesence they divide more slowly - both affecting their ability to take up DNA (transient transfection), and minimizing their abillity to incorporate the DNA into the genome (stable selection).Cultured common cell lines are often immortalized, and generally continue to aquire mutations, leading to a heterogenous population that may perform differently from cells of lower passage number - leading to results that are not reproducible.
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What is transfection efficiency?
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Transfection efficiency is a measure of how many cells take up the DNA during the transfection process. Many transfection reagents can achieve a transfection efficiency of >90% in common cell lines. Other cell lines are hard to transfect, and require special reagents and/or techniques to achieve even a small population of transfected cells.
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Why do I see a precipitate in my cell culture after lipid-based transfection?
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The precipitate is likely excess lipid or EDTA and will probablly not affect transfection efficiency. If your DNA plasmid is suspended in TE, be sure the concentration of EDTA is <0.3 mM, or suspend the DNA in sterile molecular biology grade water instead.
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