Lipo2000 Transfection Reagent
High-efficiency transfection without the need for serum-reduced medium.
One-click copy of information
Instruction manual- Product Description
- Composition
- Precautions
-
Product Storage:
Store at 4°C; shelf life: 12 months. Do not freeze!
Product Overview:
This product is a cationic liposome‑based transfection reagent that utilizes lipid nanoparticle (LNP) delivery technology, suitable for delivering nucleic acids into cultured cells. It offers exceptionally high transfection efficiency across common mammalian cell lines, excellent reproducibility, straightforward operation, and is free of visible…
It exhibits evident cytotoxicity and is suitable for both adherent and suspension cells, functioning effectively in serum‑supplemented, antibiotic‑containing complete culture media. This reagent maintains high transfection efficiency even when used with plasmids up to 13 kb in size.
Product Features
1. High transfection efficiency, simple operation, low toxicity, and excellent reproducibility. For most cell lines, such as HEK293T, HEK293F, CHO, and HeLa, transfection efficiency can reach 80%–90% when the DNA-to-transfection reagent ratio (μg:μL) is maintained at 1:2 to 1:3.
2. Excellent compatibility: transfection can be performed in the presence of serum and antibiotics, without the need to pre‑replace Opti‑MEM® or other serum‑free media.
3. The transfection reagent is a chemically synthesized product and contains no animal-derived components (Animal origin-free, AOF).
4. This product has been sterilized by filtration through a 0.22 µm membrane filter.
Instructions for DNA Transfection
(This instruction manual uses a 6-well plate and 293T cells as an example.)
1. Cell seeding
The day before transfection, seed 293T cells in a 6-well plate at 3 × 10^5 to 5 × 10^5 cells per well, and culture them in DMEM supplemented with 10% serum for 18–22 hours.
2. Transfection Process
a) Prepare DNA dilution (per well): Add 2 µg of plasmid DNA to 100 µL of serum-free DMEM culture medium and mix thoroughly.
b) Prepare the Lipo2000 dilution (per well): Take another 1.5 mL centrifuge tube and add 100 μL of serum-free DMEM culture medium and
5 µL of Lipo2000 (mix well before pipetting), gently mix, and let stand at room temperature for 5 minutes.
c) Add the DNA dilution to the Lipo2000 dilution, gently mix after combining, and incubate at room temperature for 20 minutes to form the DNA–Lipo2000 complex.
d) Using a pipette, dispense the DNA–Lipo2000 complex into the cell culture wells, and gently shake the 6-well plate to mix.
e) After 6 hours of transfection, remove the old culture medium and replace it with 2 mL of fresh DMEM containing 10% serum.
f) Continue culturing for 24 to 48 hours, then proceed with the subsequent experiments.
Reference Table for DNA and Transfection Reagent Dosages Across Different Cell Culture Vessel Sizes
Cell culture vessel
Cell culture medium volume
(per well)
Volume of serum-free culture medium for dilution DNA amount
Transfection reagent
96-well plate 100 µL 10 µL 0.1 µg 0.25 µL 48-well plate 200 µL 20 µL 0.2 µg 0.5 µL 24-well plate 500 µL 50 µL 0.5 µg 1.25 µL 12-well plate 1 ml 50 µL 1 µg 2.5 µL 6-well plate 2 ml 100 µL 2 µg 5 µL 6 cm Petri dish 4 ml 200 µL 4 µg 10 µL 10 cm Petri dish 10 ml 500 µL 10 µg 25 µL siRNA Transfection Instructions
(This protocol uses a 6-well plate and an siRNA final concentration of 50 nM as an example.)
1. Cell inoculation
The day before transfection, seed an appropriate number of cells in a 6-well plate to achieve a cell density of 30%–70% at the time of transfection.
2. Transfection Process
a) Take a sterile 1.5 mL centrifuge tube, add 100 μL of serum-free culture medium first, then add 100 pmol of siRNA, and mix well.
b) Transfer 100 μL of serum-free culture medium and 5 μL of Lipo2000 (after thoroughly mixing the solution prior to pipetting) into a separate 1.5-mL centrifuge tube. Gently mix, then incubate at room temperature for 5 minutes.
c) Add the siRNA dilution to the Lipo2000 dilution, combine the two solutions, gently mix, and incubate at room temperature for 20 minutes.
d) Use a pipette to dispense the transfection complex into the cell culture wells, and gently rock the 6-well plate to mix thoroughly.
e) After 6 hours of transfection, remove the old culture medium and replace it with 2 mL of fresh complete culture medium.
f) Continue culturing for 24 to 48 hours, and assess the expression level of the target gene using qPCR or Western blot.
Reference table for siRNA and transfection reagent amounts based on cell culture vessel size
Cell culture vessel Culture volume Volume of serum-free culture medium for dilution siRNA dosage Transfection reagent 24-hole plate 500 µL 50 µL 25 pmoles 1.5 µL 12-hole plate 1 ml 50 µL 50 pmoles 2.5 µL 6-well plate 2 ml 100 µL 100 pmoles 5 µL 6 cm Petri dish 4 ml 200 µL 200 pmoles 10 µL 10 cm Petri dish 10 ml 500 µL 500 pmoles 25 µL -
Product Composition
Item number Component Specifications 100-507 Lipo2000 transfection reagent 1.5 mL -
Precautions for DNA Transfection
1. Prior to transfection, cells must be in good growth condition. Using cells that are in the logarithmic growth phase, exhibit high viability, and have been passaged only a few times will yield better transfection results.
2. The quality of plasmid DNA is critical for successful transfection. It is recommended to use high‑purity, endotoxin‑free plasmids. The amount of plasmid DNA should be optimized according to the cell type and the specific experimental conditions.
3. Depending on the specific experimental conditions and cell type, the DNA-to-transfection reagent ratio (µg/µL) is recommended to be optimized within the range of 1:1 to 1:4. Excessive amounts may increase cytotoxicity. The transfection time should not exceed 6 hours.
4. This product is intended solely for scientific research by qualified professionals and shall not be used for clinical diagnosis or treatment, nor for food or pharmaceutical purposes, nor stored in ordinary residential premises.
5. For your safety and health, please wear a lab coat and disposable gloves while performing this procedure.
6. If you encounter any issues during use, please contact our technical support.
Precautions for siRNA Transfection
1. Prior to transfection, cells must be in good growth condition. Using cells that are in the logarithmic growth phase, exhibit high viability, and have been passaged only a few times will yield better transfection results.
2. It is recommended to use high-quality siRNA and to dissolve the lyophilized powder in RNase-free ddH2O, achieving a final concentration of 20 µM.
3. Adjust the final concentration of siRNA and the amount of transfection reagent according to the specific experimental conditions and cell type.
4. This product is intended solely for scientific research by qualified professionals and shall not be used for clinical diagnosis or treatment, nor for food or pharmaceutical purposes, nor stored in ordinary residential premises.
5. For your safety and health, please wear a lab coat and disposable gloves while performing this procedure.
6. If you encounter any issues during use, please contact our technical support.
Keywords
GOONIE
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