293 Cell-Specific Transfection Reagent is a transfection reagent that can effectively deliver DNA into 293 suspension cell lines as well as adherent cells such as HEK293T and 293FT. This product is suitable for large-scale protein production and high-efficiency viral vector expression, and has advantages including simple operation, high expression level and low cytotoxicity. It is supplied as a sterile liquid.
Experimental Materials:
• This product includes the 293 Cell-Specific Transfection Reagent.
• High-purity nucleic acid stock solution (0.5–5 μg/μL).
• Serum-free medium (e.g., O778393 Optimal-MEM Reduced Serum Medium (with Phenol Red) ).
Quality Control:
• The shelf life is 1 year when stored at 4 °C.
• Strict quality inspection has confirmed that this product is free from biological contamination.
Precautions:
• 293 Cell-Specific Transfection Reagent-nucleic acid complex at room temperature, with a total incubation time of 25 minutes. For transfection, use serum-free medium (e.g., O778393 Optimal-MEM Reduced Serum Medium (with Phenol Red)) and enzyme-free tubes.
• After transfecting cells in well plates with this product for 4–6 hours, remove the original medium and replace it with fresh serum-containing medium; when transfecting suspension cells cultured in shake flasks with this product, there is no need to replace the medium.
• This product should be stored tightly sealed at 4 °C, and repeated freezing and thawing should be avoided.
• For first-time use, the ratio of nucleic acid concentration to the 293 Cell-Specific Transfection Reagent should be optimized to achieve the highest transfection efficiency.
• For transfection, use healthy cells that are properly preserved and regularly passaged, and ensure that the medium is free from bacterial, fungal, or mycoplasma contamination. If the cells are cryopreserved cells recently recovered from liquid nitrogen, passage them at least twice before transfection.
• After using the transfection reagent, close the cap immediately to avoid prolonged exposure to air, which may affect transfection efficiency.
Example of transfection effect:
Figure 1. Cell positive rate and green fluorescent protein expression level 48 hours after delivering pDNA in Exipi293 suspension cells using the 293 Cell-Specific Transfection Reagent and competing products (24-well plate, 1 μg of EGFP-pDNA transfected per well).
Transfect cells according to the table below. The volume of each reaction mixture refers to the volume for a single well, with pipetting differences taken into account. Calculate the volumes for other wells in proportion.
293 Cell-Specific Transfection Reagent----Detailed Instructions for Transfection Steps (Taking 24-well Plate as an Example)
1. Add 1μg of DNA to 25 μL of O778393 Optimal-MEM Reduced Serum Medium (with Phenol Red) to obtain the DNA dilution.
2. Add 2 μL of 293 Cell-Specific Transfection Reagent to 25 μL of O778393 Optimal-MEM Reduced Serum Medium (with Phenol Red), and let it stand for 5 minutes. (The dosage of 293 Cell-Specific Transfection Reagent can be optimized within the range of 1–2 μL to achieve the best transfection effect.)
3. Thoroughly mix the DNA dilution from Step 1 with the diluted 293 Cell-Specific Transfection Reagent from Step 2, and incubate at room temperature for 20 minutes.
4. Mix the incubated 293 Cell-Specific Transfection Reagent-DNA complex with 450 μL of serum-free medium, then add the mixture to the cells.
5. After incubating the cells at 37°C for 4-6 hours, replace the medium with 500 μL of fresh complete medium. Continue incubating the cells at 37°C for 2-4 days before analyzing the transfected cells.
293 Cell-Specific Transfection Reagent----Detailed Instructions for Transfecting Suspension Cells (Taking a 125mL Culture Flask and 30mL Culture System as Examples)
1. Dilute 30 μg of DNA to a total volume of 300 μL using O778393 Optimal-MEM Reduced Serum Medium (with Phenol Red) to obtain the DNA dilution.
2. Add 60 μL of 293 Cell-Specific Transfection Reagent to 240 μL of O778393 Optimal-MEM Reduced Serum Medium (with Phenol Red), and let it stand for 5 minutes. (The dosage of 293 Cell-Specific Transfection Reagent can be optimized within the range of 30–60 μL to achieve the best transfection effect.)
3. Thoroughly mix the DNA dilution from Step 1 with the diluted 293 Cell-Specific Transfection Reagent from Step 2, and incubate at room temperature for 20 minutes.
4. Directly add the incubated 293 Cell-Specific Transfection Reagent-DNA complex into the cell culture flask. Follow the specific requirements in the Suspension Cell Culture Instruction Manual for the feeding protocol during the culture process. Continue incubating the cells at 37°C for 5–7 days, then analyze the cell transfection status.
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