EnoGeneFec 2000 Transfection Reagent
Cat# MBS8579497-0,5mL
Size : 0.5mL
Marca : MyBiosource
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.
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 – 1ug/ul
For Research Use Only. Not for use in diagnostic procedures. Catalog #: MBS8579497Introduction: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… More details
Product Overview
Online Datasheet in PDFProduct Name
Transfection Reagent
Full Product Name
EnoGeneFec 2000 Transfection Reagent
Concentration
1ug/ul (lot specific)
Preparation and Storage
It is stabilized for extended storage at 4 degree C for one year when very tightly closed.
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.
ISO Certification
Manufactured in an ISO 9001:2015 Certified Laboratory.
Procedures of Transfection Reagent reagent
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.
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.
Other Notes
Small volumes of Transfection Reagent reagent vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.
Searchable Terms for Transfection Reagent purchase
MBS8579497 is a reagent targeting Transfection Reagent. Also known as EnoGeneFec 2000 Transfection Reagent.