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Position：Lipo2000 transfection reagent
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Product number： DIB034-1.5ml DIB034-0.75ml
Product specifications： 1.5ml 0.75ml
Storage: Store at 2-4°C for one year.(Avoid freezing)
Product description:
Data Invention Biotech's unique patented formula Lipo2000 has a transfection efficiency between Lip2000 and Lip3000, with lower toxicity and higher transfection efficiency. It is a new type of cationic liposome transfection reagent.Suitable for transfection of nucleic acids (DNA and RNA) into eukaryotic cells, with low cytotoxicity; high transfection efficiency for various types of cells and culture plates; the presence of serum during transfection does not affect the advantages of transfection efficiency .
Scope of application: transfection of adherent cells and suspension cells (mammalian cell lines).
Transfection efficiency of common cells (for reference only, the transfection efficiency will vary with different experimental conditions)
Product parameter
Lipo2000 transfection reagent
Product number： DIB034-1.5ml DIB034-0.75ml
Product specifications： 1.5ml 0.75ml
Storage: Store at 2-4°C for one year.(Avoid freezing)
Product description:
Data Invention Biotech's unique patented formula Lipo2000 has a transfection efficiency between Lip2000 and Lip3000, with lower toxicity and higher transfection efficiency. It is a new type of cationic liposome transfection reagent.Suitable for transfection of nucleic acids (DNA and RNA) into eukaryotic cells, with low cytotoxicity; high transfection efficiency for various types of cells and culture plates; the presence of serum during transfection does not affect the advantages of transfection efficiency .
Scope of application: transfection of adherent cells and suspension cells (mammalian cell lines).
Transfection of plasmid DNA:
For most cells, the ratio of DNA (µg) to Lipo2000 (µl) is 1:2~1:3.High cell density during transfection can obtain high transfection efficiency and expression level, and can reduce cytotoxicity.
For most cells, the ratio of DNA (µg) to Lipo2000 (µl) is 1:2~1:3.High cell density during transfection can obtain high transfection efficiency and expression level, and can reduce cytotoxicity.
1. Take a 24-well plate as an example for adherent cells: On the day before transfection, inoculate 0.5-2×105 cells with 500 µl of antibiotic-free medium to reach 70-90% confluence the next day.Suspension cells: Before preparing the DNA-Lip2000 complex, inoculate 4-8×105 cells with 500 µl antibiotic-free medium.
2. For each transfection sample, perform the following operations: a. Add 50 µl Opti-MEM I ReLipced Serum Medium and 0.8 µg DNA to the eppendorf tube and mix gently to make a DNA dilution.b. Add 50 µl Opti-MEMI ReLipced Serum Medium and 2.0 µl Lipo2000 to another eppendorf tube (note that you should mix well before use), mix gently to make a Lip2000 dilution, and let it stand at room temperature for 5 minutes.c. Mix the DNA diluent and Lip2000 diluent, mix gently, and let stand at room temperature for 20 minutes to form a DNA-Lip2000 complex.The DNA-Lip2000 complex can exist stably for 6 hours at room temperature.
3. Add the DNA-Lip2000 complex to the inoculated cells, and gently shake the culture plate back and forth to make the complex evenly dispersed.
4. After culturing for 4-6 hours in a 37°C CO2 incubator, change the medium and continue culturing for 18 to 48 hours.
5. If you want to screen for stable cell lines, inoculate the cells in a fresh medium at a ratio of 1:10 or higher 24 hours after transfection, and add selective medium for screening the next day.
2. For each transfection sample, perform the following operations: a. Add 50 µl Opti-MEM I ReLipced Serum Medium and 0.8 µg DNA to the eppendorf tube and mix gently to make a DNA dilution.b. Add 50 µl Opti-MEMI ReLipced Serum Medium and 2.0 µl Lipo2000 to another eppendorf tube (note that you should mix well before use), mix gently to make a Lip2000 dilution, and let it stand at room temperature for 5 minutes.c. Mix the DNA diluent and Lip2000 diluent, mix gently, and let stand at room temperature for 20 minutes to form a DNA-Lip2000 complex.The DNA-Lip2000 complex can exist stably for 6 hours at room temperature.
3. Add the DNA-Lip2000 complex to the inoculated cells, and gently shake the culture plate back and forth to make the complex evenly dispersed.
4. After culturing for 4-6 hours in a 37°C CO2 incubator, change the medium and continue culturing for 18 to 48 hours.
5. If you want to screen for stable cell lines, inoculate the cells in a fresh medium at a ratio of 1:10 or higher 24 hours after transfection, and add selective medium for screening the next day.
Optimization of plasmid DNA transfection In order to achieve the highest transfection efficiency and reduce the influence of cytotoxicity, the ratio of DNA to Lip2000 and cell density can be optimized. Generally, DNA (µg) is optimized in the range of 1:0.5~1:5 And Lip2000 (µl) ratio.
The amount of media, nucleic acid and Lipo2000 for transfection in different cell culture plates
The amount of media, nucleic acid and Lipo2000 for transfection in different cell culture plates
| Cell culture plate | Area per hole | Medium amount | DNA Transfection | siRNA | |||||||
| The amount of plating medium | The amount of diluted medium | ||||||||||
| 96-well | 0.3cm2 | 100ul | 2*25ul | 0.2ug | 0.5ul | 5pmol | 0.25ul | ||||
| 24-well | 2cm2 | 500ul | 2*50ul | 0.8ug | 2ul | 20pmol | 1.0ul | ||||
| 12-well | 4cm2 | 1ml | 2*100ul | 1.6ug | 4ul | 40pmol | 2.0ul | ||||
| 6-well | 10cm2 | 2ml | 2*250ul | 4ug | 10ul | 100pmol | 5ul | ||||
| 60-mm | 20cm2 | 5ml | 2*0.5ml | 8ug | 20ul | 200pmol | 10ul | ||||
| 10-cm | 60cm2 | 15ml | 2*1.5ml | 24ug | 60ul | 600pmol | 30ul | ||||
Transfection efficiency of common cells (for reference only, the transfection efficiency will vary with different experimental conditions)
| Cell type | HEK293 | HCT116 | WRL-68 | HepG2 | NH/3T3 | THP-1 | Hela | MCF-7 | 293T | TS cell | HO1980 | A549 |
| Transfection efficiency | ＞80% | ＞80% | ~80% | ~80% | ~80% | ＞50% | ＞80% | ＞80% | ＞80% | ＞60% | ＞60% | ＞80% |
| Cell type | MEF | Chok1 | Hep3B | C2C12 | Neuro-2a | HUVEC | MDCK | Hep2C | WHI | B50 | Calu1 | L929 |
| Transfection efficiency | ＞50% | ＞50% | ＞80% | ＞80% | ＞70% | ＞80% | ＞80% | ＞80% | ＞80% | ＞70% | ＞70% | ＞70% |
Product parameter
| Name | Lipo2000transfection reagent | ||||||||||
| CAT# | DIB034-1.5ml DIB034-0.75ml | ||||||||||
| CAS# | N/A | ||||||||||
| Storage# | 4℃ dry and avoid light | ||||||||||
| Shelf Life# | 12 months | ||||||||||
| Ex(nm)# | N/A | ||||||||||
| Em(nm)# | N/A | ||||||||||
| MW# | N/A | ||||||||||
| Solvent# | N/A | ||||||||||