AffiGEN® Transfection Reagent

AffiGEN® Transfection Reagent

AffiGEN® Transfection Reagent is a liposome-mediated transfection reagent. It is applicable for plasmid DNA and RNA transfection of various adherent or suspension cells. AffiGEN® Transfection Reagent has the excellent ability to form liposome / nucleic acid complexes and rapid release of nucleic acid transfected into cells, which ensures excellent transfection performance and extremely low cytotoxicity. The presence of serum and antibiotics does not affect its transfection effect, so the formed liposome/nucleic acid complex can be directly added to the complete medium. There is no need to change the medium before and after transfection, which simplifies the operation.

Components:

Storage:

Store at 2 ~ 8°C. Adjust the shipping method according to the destination.

Please mix this product up and down before use, do not freeze.

Applications:

It is applicable for most eukaryotic cells: 293T, A549, HeLa, SW480, 4T1, Jurkat, NIH3T3, MDA-MB-453, Vero, K562, Raw264.7, etc.

Notes:

For research use only. Not for use in diagnostic procedures.

1. Cell density varies greatly with cell lines, and cell density directly determines transfection efficiency. In general, transfection can be performed when the cell density reaches 60% - 80% to achieve higher transfection efficiency. However, the optimal transfection density
2. varies from cell to cell. Therefore, when transfecting a specific cell for the first time, the optimal transfection density of the cell can be confirmed by pre-experiment. In addition, please try to keep the same inoculum ratio during the transfection process to improve the repeatability of the experiment.
3. Guarantee high-quality DNA of high purity, sterility, and endotoxin-free.
4. The ratio of AffiGEN® Transfection Reagent to DNA: When transfecting certain cells for the first time, the recommended ratio of DNA to AffiGEN® Transfection Reagent is 1:3. That is, 3 μL AffiGEN® Transfection Reagent is used for 1 μg DNA. The usage of AffiGEN® Transfection Reagent used to transfect 1 μg of DNA can be adjusted between 1 - 5 μL to obtain better transfection efficiency.
5. Cell transfection experiments should be performed gently. Note that AffiGEN® Transfection Reagent and DNA should be diluted separately with opti-MEM first, then add DNA/opti-MEM dropwise to AffiGEN® Transfection Reagent/opti-MEM. Mix well to form complexes.

Experiment Process:

Transient transfection of cells (take 24-well plate transfection as an example).

• Cell inoculation:

1. Subculture the cells about 24 hours before transfection, and inoculate at a density of about 0.5 - 2 × 10^5 cells/well.
2. Overnight incubation.

• Formation of AffiGEN® Transfection Reagent/DNA complexs:

1. Add 25 μL opti-MEM medium and 1.5 μL AffiGEN® Transfection Reagent to a 1.5 mL sterile centrifuge tube, then mix gently with a pipette.
2. Add 25 μL opti-MEM medium and 0.5 μg DNA to a 1.5 mL sterile centrifuge tube, then mix gently with a pipette.
3. Add DNA/opti-MEM to AffiGEN® Transfection Reagent/opti-MEM, mix gently with a pipette, and let it stand at room temperature for 5 min before transfection.

• Transfection:

1. Add the AffiGEN® Transfection Reagent/DNA complex mixture to the culture medium, then shake the petri dish gently to disperse it evenly. For special cases, the fresh serum-containing medium can be replaced before transfection to prevent cell death caused by excessive cell density and insufficient nutrition during the culture period.
2. Overnight incubation for 24 - 48 h. For special cases, the fresh serum-containing medium can be replaced before transfection to prevent cell death caused by excessive cell density and insufficient nutrition during the culture period.

Recommended transfection system:

The above protocol is for reference only and can be optimized according to the specific situation.