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.
L3287
Escort™ IV Transfection Reagent
Lipid reagent for transient and stable transfection of mammalian and insect cells.
Synonym(s):
Gene delivery
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About This Item
UNSPSC Code:
41106502
NACRES:
NA.25
Skip To
grade
Molecular Biology
Quality Level
form
liquid (aqueous solution)
usage
1 mL sufficient for 160-500 transfections
concentration
1 mg/mL
technique(s)
transfection: suitable
storage temp.
2-8°C
Related Categories
1 of 4
This Item | L3037 | XTGHP-RO | 11202375001 |
|---|---|---|---|
| grade for molecular biology | grade for molecular biology | grade for molecular biology | grade - |
| technique(s) transfection: suitable | technique(s) transfection: suitable | technique(s) transfection: suitable | technique(s) transfection: suitable |
| form liquid (aqueous solution) | form liquid (aqueous solution) | form liquid (aqueous solution) | form liquid |
| concentration 1 mg/mL | concentration 1 mg/mL | concentration - | concentration - |
| Quality Level 100 | Quality Level 200 | Quality Level 100 | Quality Level 100 |
| storage temp. 2-8°C | storage temp. 2-8°C | storage temp. −20°C | storage temp. 2-8°C |
General description
Escort™ IV 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 a wide variety of eukaryotic cell types.
Application
Suitable for transient and stable transfection of nucleic acids into cultured eukaryotic cells. Use approximately 4-16 μL Escort™ IV and 2 μg DNA per 6 cm cell culture plate. Protocol optimization provides very efficient transfection. For a list of cells that have been successfully transfected using Escort™ IV, see the Transfection Reagent Selection Guide.
Biochem/physiol Actions
A stable complex is formed when Escort™ IV 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 cell lines
- Low toxicity
- Compatible with both serum and serum-free transfection protocols
- Ideal for Sf9, Sf21 and S2 insect cells
Other Notes
Escort™ IV formulation:
1 mg/mL total lipid in water
Note the identity of the lipids used in Escort™ IV is confidential.
1 mg/mL total lipid in water
Note the identity of the lipids used in Escort™ IV is confidential.
Legal Information
Escort is a trademark of Sigma-Aldrich Co. LLC
Disclaimer
Do not freeze.
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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Victoria J Hammond et al.
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Sol-gel nanoprobes, also known as Photonic Explorer for Bioanalysis with Biologically Localised Embedding (PEBBLE), capable of performing in-vitro intracellular monitoring of reactive oxygen species have been developed using a modified form of 5(6)-carboxyfluorescein diacetate. A sol-gel matrix was selected for
R'ada Massarwa et al.
Developmental cell, 12(4), 557-569 (2007-04-11)
Formation of syncytial muscle fibers involves repeated rounds of cell fusion between growing myotubes and neighboring myoblasts. We have established that Wsp, the Drosophila homolog of the WASp family of microfilament nucleation-promoting factors, is an essential facilitator of myoblast fusion
Eva Krieghoff et al.
Journal of cell science, 119(Pt 7), 1453-1463 (2006-03-24)
beta-catenin is the central signalling molecule of the canonical Wnt pathway, where it activates target genes in a complex with LEF/TCF transcription factors in the nucleus. The regulation of beta-catenin activity is thought to occur mainly on the level of
Zev Bryant et al.
Proceedings of the National Academy of Sciences of the United States of America, 104(3), 772-777 (2006-12-22)
Myosin VI supports movement toward the (-) end of actin filaments, despite sharing extensive sequence and structural homology with (+)-end-directed myosins. A class-specific stretch of amino acids inserted between the converter domain and the lever arm was proposed to provide
Manveen K Gupta et al.
Antioxidants & redox signaling, 8(5-6), 1081-1093 (2006-06-15)
Cardiac myocytes, upon exposure to increasing doses of norepinephrine (NE), transit from hypertrophic to apoptotic phenotype. Since reactive oxygen species (ROS) generation is attributed to both phenomena, the authors tested whether an elevation in intracellular ROS level causes such transition.
Articles
This brief webinar provides an overview of what transfection is and the methods that are used to introduce DNA or RNA into eukaryotic cells.
Transfection introduces genetic material into cells, aiding research in gene expression and cell biology.
Protocols
Universal Transfection Reagent enables efficient nucleic acid delivery into various cells, compatible with different cell culture conditions.
Incorporation of DNA into eukaryotic cells using calcium phosphate is a common method of transfection. This transfection protocol can be optimized for a wide variety of cell types.
The product bulletin providin detailed use protocol for easy DNA transfection.
Using liposomes for transfection is a method for introducing DNA into eukaryotic cells. Escort™ IV Transfection Reagent is composed of a polycationic lipid and a neutral, non-transfecting lipid compound.
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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How can I increase the efficiency of my transfection?
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What quality does the DNA need to be in order to use it for transfection?
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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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What is the Department of Transportation shipping information for this product?
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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.
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How do I choose a transfection reagent?
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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.
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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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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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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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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
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Can antibiotics be present in the medium during transfection?
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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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