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.
L3037
Escort™ III Transfection Reagent
Lipid reagent for transfecting sensitive and primary cells
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About This Item
UNSPSC Code:
41106502
NACRES:
NA.25
Pricing and availability is not currently available.
Recommended Products
grade
for molecular biology
Quality Level
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)
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
Components
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.
Caution
Do not freeze.
Principle
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.
Legal Information
Escort is a trademark of Sigma-Aldrich Co. LLC
related product
Product No.
Description
Pricing
Storage Class Code
10 - Combustible liquids
WGK
WGK 3
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Personal Protective Equipment
dust mask type N95 (US), Eyeshields, Gloves
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Patrick Orth et al.
Molecular biotechnology, 38(2), 137-144 (2008-01-26)
The aim of the present study was to evaluate the efficacy of novel nonviral gene delivery systems in cells of musculoskeletal origin. Primary cultures of lapine skeletal muscle cells, lapine articular chondrocytes, human cells from fibrous dysplasia and cell lines
Tomás C O'Riordan et al.
American journal of physiology. Regulatory, integrative and comparative physiology, 292(4), R1613-R1620 (2006-12-16)
The development and application of a methodology for measurement of oxygen within single mammalian cells are presented, which employ novel macromolecular near infrared (NIR) oxygen probes based on new metalloporphyrin dyes. The probes, which display optimal spectral characteristics and sensitivity
Lixian Liu et al.
Frontiers in cell and developmental biology, 9, 634242-634242 (2021-03-12)
The mitogen-inducible gene 6 (MIG6) is an adaptor protein widely expressed in vascular endothelial cells. However, it remains unknown thus far whether it plays a role in angiogenesis. Here, using comprehensive in vitro and in vivo model systems, we unveil
Inmaculada Navarro-Lérida et al.
Journal of cell science, 117(Pt 9), 1687-1697 (2004-04-13)
Several cell types express inducible nitric oxide synthase (NOS2) in response to exogenous insults such as bacterial lipopolysaccharide (LPS) or proinflammatory cytokines. For instance, muscular cells treated with LPS and interferon gamma (IFN-gamma) respond by increasing the mRNA and protein
Thomas Iskratsch et al.
The Journal of cell biology, 191(6), 1159-1172 (2010-12-15)
Members of the formin family are important for actin filament nucleation and elongation. We have identified a novel striated muscle-specific splice variant of the formin FHOD3 that introduces a casein kinase 2 (CK2) phosphorylation site. The specific targeting of muscle
Articles
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.
Protocols
The product bulletin providin detailed use protocol for easy DNA transfection.
Product manual provides detailed protocol for easy DNA transfection.
Related Content
Browse our convenient transfection reagent selection guide to match the best reagent for your specific cell line and application needs.
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Can I transfect cells plated at low density?
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What is the Department of Transportation shipping information for this product?
1 answer-
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
Helpful?
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Is low cell passage number an important consideration for transfection?
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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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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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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 increase the efficiency of my transfection?
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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.
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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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Which Escort™ transfection reagent should I use for my application?
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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.
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How can I determine the efficiency of my transfection?
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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
Helpful?
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Is the size of the plasmid an important consideration for transfection?
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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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