Description:GenMute™ siRNA Transfection Reagent for HepG2 Cell is pre-optimized for transfecting siRNA to HepG2 cells. HepG2 (Hepatocellular carcinoma, human) is a perpetual add cell line which was derived from the liver tissue of a 15 year old caucasian male with a well differentiated hepatocellular carcinoma. These cells are epithelial in morphology, have a model chromosome number of 55 and are not tumorigenic in nude mice. The cells secrete a variety of major plasma proteins e.g.. albumin, alpha 2-macroglobulin, alpha 1-antitrypsin, transferrin and plasminogen. They have been grown successfully in large scale cultivation systems. Hepatitis B virus surface antigens have not been detected. HepG2 cells have been shown to be G418 resistant (400µg/mL). The cells will respond to stimulation with human growth hormone. Refer to the following optimal transfection conditions for maximal silencing on HepG2 cells. GenMute™ reagent, 1.0 ml, is sufficient for ~833 transfections in 24 well plates or ~416 transfections in 6 well plates.

Summary of Optimal Transfection Conditions:Cell culture mediumConfluency of cell on the day of transfectionOptimal siRNA concentrationDiluent for siRNA and GenMute™ ReagentIncubation Time to Form GenMute™/siRNA ComplexMaximal Silencing Efficiency Transfection Results:Reporter GeneSilencing Efficiency (% )

DMEM with 4.5 g/L glucose, 10% FBS~50%40 nM1xGenMute™ Transfection Buffer~15 minutes at RT48 hoursRenilla Luciferase85%

Storage:Store at 4 °C. If stored properly, the product is stable for 12 months or longer.

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