Application Data
(1. 18 to 24 hours prior to transfection, trypsinize and adjust the cell concentration, seed cells at a density of 1-3 x 105 cells per well in 2.0ml of appropriate growth medium (with serum and antibiotics if cells are cultured in the presence of them). Incubate the cells at 37 degree C in a CO2 incubator until cells are 70% to 90% confluent at the time of transfection.2. For each transfection sample, prepare the complexes as follows:Solution A: Dilute 2.0 ug of DNA into 100 ul of serum-free, antibiotic-free medium.Solution B: Vortex EnoGeneFec™ 2000 Transfection Reagent thoroughly prior use, then dilute 10 ul of EnoGeneFec™ 2000 Transfection Reagent in 100 ul serum-free, antibiotic-free medium. Incubate Solution A and B at room temperature for 5 minutes.3. Combine the solutions, mix gently to ensure uniform distribution and incubate for 20 minutes at room temperature.NOTE: Complexes are stable at room temperature for 3-5 hours.4. Add 0.8 ml of serum-free, antibiotic-free medium to EnoGeneFec™-DNA complex. Mix solution gently.5. Remove growth medium from the cells and add 1.0 ml of EnoGeneFec™-DNA solution to the each well containing cells.6. After 5-8 hours, remove transfection solution and add 2.0 ml of the appropriate growth medium (with serum and antibiotics). Incubate the cells at 37 degree C in a CO2 incubator for a total of 24-72 hours.7. Assay cell extracts or phenotype for gene activity 24-72 hours after the start of transfection depending on the cell types and promoter activity. To make stable cell lines: Passage cells at a 1:10 (or higher dilution) into fresh growth medium 24 hours post transfection. Selection medium can be added the following day if desired.Use the following conditions as guidelines to transfect adherent mammalian cells in a 6-well or 35mm dish format. For other culture vessels, please refer to Table 1.)
Transfection Reagent
EnoGeneFec 2000 Transfection Reagent
Synonyms
Transfection Reagent; N/A; EnoGeneFec 2000 Transfection Reagent; Transfection Reagent reagent
Concentration
1ug/ul (varies by lot)
Important Notes
Required amount of EnoGeneFec™ 2000 Transfection Reagent
For initial optimization experiments, transfect a monolayer of cells that is 70–90% confluent, using 2:1~ 5:1 ratios of EnoGeneFec™ 2000 Transfection Reagent (ul) to DNA (ug), respectively. For most cell types, the EnoGeneFec™ 2000 Transfection Reagent: DNA (2:1) ratio provides excellent transfection level. For some hard-to-transfect cells, the EnoGeneFec™ 2000 Transfection Reagent: DNA (5:1) ratio is recommended.
For initial optimization experiments, transfect a monolayer of cells that is 70–90% confluent, using 2:1~ 5:1 ratios of EnoGeneFec™ 2000 Transfection Reagent (ul) to DNA (ug), respectively. For most cell types, the EnoGeneFec™ 2000 Transfection Reagent: DNA (2:1) ratio provides excellent transfection level. For some hard-to-transfect cells, the EnoGeneFec™ 2000 Transfection Reagent: DNA (5:1) ratio is recommended.
Cell culture
A healthy cell culture lays the foundation for successful transfection. Different cells or cell lines have very specific media, serum, and supplement requirements. Low passage number (<50 splitting cycles) ensures that the cell genotype has not become altered. We recommend subculturing cells 24 hours before transfection. This provides normal cell metabolism and increases the likelihood of DNA uptake.
Effect of serum
We do not recommend using serum during complex formation between EnoGeneFec™ 2000 Transfection Reagent and plasmid DNA, as serum may inhibit complex formation.
Plasmid DNA quality
It is critical to accurately determine the plasmid DNA concentration using 260nm absorption. DNA content must be determined by 260nm absorption (estimates of DNA content based on the intensity of gel bands are not sufficiently accurate). Determine the DNA purity using a 260 nm/280 nm ratio; the ratio should be 1.8.
Preparation and Storage
It is stabilized for extended storage at 4 degree C for one year when very tightly closed.
Application Data
(1. 18 to 24 hours prior to transfection, trypsinize and adjust the cell concentration, seed cells at a density of 1-3 x 105 cells per well in 2.0ml of appropriate growth medium (with serum and antibiotics if cells are cultured in the presence of them). Incubate the cells at 37 degree C in a CO2 incubator until cells are 70% to 90% confluent at the time of transfection.2. For each transfection sample, prepare the complexes as follows:Solution A: Dilute 2.0 ug of DNA into 100 ul of serum-free, antibiotic-free medium.Solution B: Vortex EnoGeneFec™ 2000 Transfection Reagent thoroughly prior use, then dilute 10 ul of EnoGeneFec™ 2000 Transfection Reagent in 100 ul serum-free, antibiotic-free medium. Incubate Solution A and B at room temperature for 5 minutes.3. Combine the solutions, mix gently to ensure uniform distribution and incubate for 20 minutes at room temperature.NOTE: Complexes are stable at room temperature for 3-5 hours.4. Add 0.8 ml of serum-free, antibiotic-free medium to EnoGeneFec™-DNA complex. Mix solution gently.5. Remove growth medium from the cells and add 1.0 ml of EnoGeneFec™-DNA solution to the each well containing cells.6. After 5-8 hours, remove transfection solution and add 2.0 ml of the appropriate growth medium (with serum and antibiotics). Incubate the cells at 37 degree C in a CO2 incubator for a total of 24-72 hours.7. Assay cell extracts or phenotype for gene activity 24-72 hours after the start of transfection depending on the cell types and promoter activity. To make stable cell lines: Passage cells at a 1:10 (or higher dilution) into fresh growth medium 24 hours post transfection. Selection medium can be added the following day if desired.Use the following conditions as guidelines to transfect adherent mammalian cells in a 6-well or 35mm dish format. For other culture vessels, please refer to Table 1.)
Application Data
(1. 18 to 24 hours prior to transfection, trypsinize and adjust the cell concentration, seed cells at a density of 1-3 x 105 cells per well in 2.0ml of appropriate growth medium (with serum and antibiotics if cells are cultured in the presence of them). Incubate the cells at 37 degree C in a CO2 incubator until cells are 70% to 90% confluent at the time of transfection.2. For each transfection sample, prepare the complexes as follows:Solution A: Dilute 2.0 ug of DNA into 100 ul of serum-free, antibiotic-free medium.Solution B: Vortex EnoGeneFec™ 2000 Transfection Reagent thoroughly prior use, then dilute 10 ul of EnoGeneFec™ 2000 Transfection Reagent in 100 ul serum-free, antibiotic-free medium. Incubate Solution A and B at room temperature for 5 minutes.3. Combine the solutions, mix gently to ensure uniform distribution and incubate for 20 minutes at room temperature.NOTE: Complexes are stable at room temperature for 3-5 hours.4. Add 0.8 ml of serum-free, antibiotic-free medium to EnoGeneFec™-DNA complex. Mix solution gently.5. Remove growth medium from the cells and add 1.0 ml of EnoGeneFec™-DNA solution to the each well containing cells.6. After 5-8 hours, remove transfection solution and add 2.0 ml of the appropriate growth medium (with serum and antibiotics). Incubate the cells at 37 degree C in a CO2 incubator for a total of 24-72 hours.7. Assay cell extracts or phenotype for gene activity 24-72 hours after the start of transfection depending on the cell types and promoter activity. To make stable cell lines: Passage cells at a 1:10 (or higher dilution) into fresh growth medium 24 hours post transfection. Selection medium can be added the following day if desired.Use the following conditions as guidelines to transfect adherent mammalian cells in a 6-well or 35mm dish format. For other culture vessels, please refer to Table 1.)
Related Product Information for Transfection Reagent reagent
Introduction:
EnoGeneFec™ 2000 Transfection Reagent comprises of unique formulations of polycations and liposomes representing a new class of transfection reagent designed for adherent mammalian cells to get outstanding transfection results, which will guarantee higher transfection efficiency and lower cytotoxicity for almost all adherent cell lines. EnoGeneFec™ 2000 Transfection Reagent offers significant advantages over many other transfection methods.
EnoGeneFec™ 2000 Transfection Reagent offers:
- High transfection efficiency of plasmids, antisense oligonucleotides, siRNA in almost all adherent cell types
- Fast procedure — transfection-complex formation in just 20 minutes
- Decreased cytotoxicity
EnoGeneFec™ 2000 Transfection Reagent comprises of unique formulations of polycations and liposomes representing a new class of transfection reagent designed for adherent mammalian cells to get outstanding transfection results, which will guarantee higher transfection efficiency and lower cytotoxicity for almost all adherent cell lines. EnoGeneFec™ 2000 Transfection Reagent offers significant advantages over many other transfection methods.
EnoGeneFec™ 2000 Transfection Reagent offers:
- High transfection efficiency of plasmids, antisense oligonucleotides, siRNA in almost all adherent cell types
- Fast procedure — transfection-complex formation in just 20 minutes
- Decreased cytotoxicity
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Product Notes
The Transfection Reagent (Catalog #AAA269062) is a Reagent and is intended for research purposes only. The product is available for immediate purchase. It is sometimes possible for the material contained within the vial of "Transfection Reagent, Reagent" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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