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.
L3037
Escort™ III Transfection Reagent
Lipid reagent for transfecting sensitive and primary cells
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| Size/SKU | Availability | Price |
|---|---|---|
1 mL | Check Cart for Availability | RM 1,393.00 |
About This Item
NACRES:
NA.25
UNSPSC Code:
41106502
Form:
liquid (aqueous solution)
Grade:
Molecular Biology
Technique(s):
transfection: suitable
Concentration:
1 mg/mL
RM 1,393.00
Check Cart for Availability
grade
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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This Item | |
|---|---|
| description Lipid reagent for transfecting sensitive and primary cells | description Lipid reagent for transient and stable transfection of mammalian and insect cells. |
| grade Molecular Biology | grade Molecular Biology |
| technique(s) transfection: suitable | technique(s) transfection: suitable |
| form liquid (aqueous solution) | form liquid (aqueous solution) |
| Quality Level 200 | Quality Level 100 |
| concentration 1 mg/mL | concentration 1 mg/mL |
| storage temp. 2-8°C | storage temp. 2-8°C |
Storage Class
10 - Combustible liquids
WGK
WGK 3
Flash Point (°F)
Not applicable
Flash Point (°C)
Not applicable
PPE (personal protective equipment)
Eyeshields, Gloves
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Is optimizing the transfection protocol important?
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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.
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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 are the differences between the two Escort™ products?
1 answer-
Each Escort™ transfection Reagent is a different lipid formulation. These different formulations are more readily taken up by different cells, presumably by endocytosis.Escort™ III is a unique formulation of a proprietary polycationic lipid and a neutral, non-transfecting lipid.Escort™ IV is a a confidential lipid.
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.
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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.
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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.
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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.
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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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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.
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