Cat. No. FT101

Ordering Information:

Storage: at 2-8oC for one year (avoid freezing)

Description

TransLipid® Transfection Reagent is a proprietary formulated cationic lipid that offers superior transfection efficiency across a broad range of mammalian cell lines.

• Transfect DNA and/or RNA.

• Adherent or suspension cells.

• Can be used in the presence of serum.

Applications

Transfection of adherent cells and suspension cells (mammalian cell lines)

Features

• High efficiency, low toxicity

• High loading efficiency to nucleic acid

• High affinity to cell membrane

• High stability in vitro

For most cells, it is suggested that ratio of DNA to translipid is 1:2-1:3. To obtain better transfection efficiency, we suggest to use high density cell (70-90% confluency).

Plasmid DNA Transfection

Use 24-well format as example, the experimental procedure is listed as follows

1. Inoculate cells: inoculate 0.5-2×105 cells in each well to make cells confluency up to 70-90% at the time of transfection

2. Grow cells for 12-24 hours

3. Dilute plasmid: dilute 0.8 μg plasmid with 50 μl Opti-MEM medium

Dilute TransLipid: dilute 2 μl TransLipid with 50 μl Opti-MEM medium

4. Incubate for 5 minutes at room temperature.

5. Gently mix DNA and TransLipid

6. Incubate for 20 hours at room temperature

7. Add the DNA- TransLipid complexes to each well containing cells and medium, incubate cells at 37oC in a CO2 incubator

8. Change medium after 4-6 hours transfection, then grow cells for 18-72 hours

siRNA Transfection

Take 24-well format as example, cells should be 30-50% conflueny at the time of transfection. The amount of siRNA and TransLipid required for transfection is 20 pmol and 1 μl respectively. The experimental procedure is the same as DNA transfection listed above.

Optimizing Plasmid DNA Transfection

In order to achieve the maximum transfection efficiency and minimize the effect of cytotoxicity, the ratio of DNA or siRNA to TransLipid as well as the initial cell density for transfection could be optimized. DNA transfection can be optimized within the range of 1:2-1:5, it is recommended to use a range of 10 to 50 pmol of siRNA and 0.5 to 1.5 μl of TransLipid

CITATIONS

Li J, et al. 2013. Autophagy promotes hepatocellular carcinoma cell invasion through activation of epithelial-mesenchymal transition. 34(6):1343-51. Carcinogenesis.IF=5.635. PMID: 23430956

Xu G, et al. 2013. MicroRNA-21 promotes hepatocellular carcinoma HepG2 cell proliferation through repression of mitogen-activated protein kinase-kinase 3. 13(1):469. BMC Cancer.IF=3.333. PMID: 24112539.

Xu G, et al. 2013. microR-142-3p Down-regulates IRAK-1 In Response to Mycobacterium bovis BCG Infection in Macrophages. pii: S1472-9792(13)00155-8. Tuberculosis. IF=3.033. PMID: 24053976.

Wei J, et al. 2013. MyD88 as a target of microRNA-203 in regulation of lipopolysaccharide or Bacille Calmette-Guerin induced inflammatory response of macrophage RAW264. 7 cells. 55(3-4):303-9. Mol Immunol.IF=2.645. PMID: 23522925.

Duan J, et al. 2012. PKHD1 post-transcriptionally modulated by miR-365-1 inhibits cell-cell adhesion. 30(5):382-9. Cell Biochem Funct.IF=1.854. PMID: 22411058.

He J, et al. 2012. Expression pattern of adipocyte fatty acid-binding protein gene in different tissues and its regulation of genes related to adipocyte differentiation in duck. 91(9):2270-4. Poult Sci.IF=1.516. PMID: 22912462.