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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L3037
Escort™ III Transfection Reagent
Lipid reagent for transfecting sensitive and primary cells
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| Ihnen/SKU | Verfügbarkeit | Preis |
|---|---|---|
1 mL | Warenkorb auf Verfügbarkeit prüfen | € 266,00 |
Über diesen Artikel
NACRES:
NA.25
UNSPSC Code:
41106502
Form:
liquid (aqueous solution)
Grade:
Molecular Biology
Technique(s):
transfection: suitable
Concentration:
1 mg/mL
€ 266,00
Warenkorb auf Verfügbarkeit prüfen
Technischer Dienst
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Unterstützung erhaltengrade
Molecular Biology
Quality Segment
form
liquid (aqueous solution)
usage
1 mL sufficient for 250-1000 transfections
concentration
1 mg/mL
technique(s)
transfection: suitable
storage temp.
2-8°C
General description
Escort™ III is a unique formulation of a proprietary polycationic lipid and a neutral non-transfecting lipid. This liposome-forming compound is used for transfection of nucleic acids into primary cells.
Application
Suitable for transient and stable transfection of nucleic acids into cultured eukaryotic cells. Use approximately 2-8 μl Escort™ III and 2 μg DNA per 6 cm cell culture plate. Protocol optimization provides very efficient transfection. The following cells have been successfully transfected using Escort™ III:
A549
C2C12 myotubes
Cardiomyocytes (rat)
COS-7
Fibroblasts (rat)
Germ cells (male rat)
Hepatocytes (rat and hamster)
HepG2
HeLa
Jurkat
Keratinocytes (human)
Myoblasts (mouse and quail)
Myocytes (mouse)
NIH3T3
PC-12
Retinal Neurons (rat)
Tracheobronchial cells (sheep)
A549
C2C12 myotubes
Cardiomyocytes (rat)
COS-7
Fibroblasts (rat)
Germ cells (male rat)
Hepatocytes (rat and hamster)
HepG2
HeLa
Jurkat
Keratinocytes (human)
Myoblasts (mouse and quail)
Myocytes (mouse)
NIH3T3
PC-12
Retinal Neurons (rat)
Tracheobronchial cells (sheep)
Biochem/physiol Actions
A stable complex is formed when Escort™ III 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
- Suitable for stable and transient transfection
- Optimized for a wide variety of primary cells
- Low toxicity
- Compatible with both serum and serum-free transfection protocols
- Ideal for PC-12 cells
Other Notes
Escort™ III formulation:
1 mg/mL total lipid in water
Note the identity of the lipids used in Escort™ III is confidential.
1 mg/mL total lipid in water
Note the identity of the lipids used in Escort™ III is confidential.
Legal Information
Escort is a trademark of Sigma-Aldrich Co. LLC
Disclaimer
Do not freeze.
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Dieser Artikel | |||
|---|---|---|---|
| form liquid (aqueous solution) | form liquid (aqueous 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 |
| concentration 1 mg/mL | concentration 1 mg/mL | concentration - | concentration - |
| Quality Level 200 | Quality Level 100 | Quality Level 200 | Quality Level - |
| storage temp. 2-8°C | storage temp. 2-8°C | storage temp. 2-8°C | storage temp. 2-8°C |
Lagerklasse
10 - Combustible liquids
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Gloves
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Is optimizing the transfection protocol important?
1 answer-
Helpful?
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Which Escort™ transfection reagent should I use for my application?
1 answer-
Escort™ III is best for sensitive cells and primary cells, particularly PC-12 and Jurkat.Escort™ IV is excellent for mammalian cell lines, primary cells and insect cells.
Helpful?
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Why do I see a precipitate in my cell culture after lipid-based transfection?
1 answer-
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.
Helpful?
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What is the difference between stable and transient transfection?
1 answer-
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.
Helpful?
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Can I transfect cells plated at low density?
1 answer-
For most transfections, cells should be >70% confluency the day of transfection, and growing in mid-log phase. Some transfection reagents are now designed to work with cells at low density, when required.
Helpful?
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How do I choose a transfection reagent?
1 answer-
There are many guides that help you select a transfection reagent. In general, consider:The type of cell(s) you will transfectThe type of nucleic acid or protein you will introduce to the cellThe composition of your cell culture mediumThe need for stable or transient transfectionThe equipment you have availableThe other factors important to you - cost, protocol flexibility, ease of use, etc.
Helpful?
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How can I increase the efficiency of my transfection?
1 answer-
Transfection efficiency is affected by many different things, including plasmid size and purity, media components present, transfection reagent selected, amount of DNA and transfection reagent used, cell density, etc. Optimizing the protocol with respect to these concerns will allow you to achieve a higher transfection efficiency. For many cell lines and transfection reagents, optimized protocols are already available.
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.
Helpful?
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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.
Helpful?
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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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