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PROTEOJUICE PROTEIN TRANSFECTION RE
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PolyJet™ DNA转
Description
GenJet™ DNA In Vitro Tranfection Reagent for HepG2 Cells is pre-optimized for HepG2 cell transfection. 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 transfection efficiency on HepG2 cells. GenJet™ reagent, 1.0 ml, is sufficient for 300 to 600 transfections in 24 well plates or 150 to 300 transfections in 6 well plates.
| Summary of Optimal Transfection Conditions: HepG2 cell culture Confluence on the day of transfection Cell culture conditions GenJet™ (µl) : DNA (µg) Ratio Diluent for DNA and Transfection Reagent Incubation Time to Form GenJet™/DNA Complex Presence of Serum/Antibiotics during Transfection Change Medium 5 Hours After Transfection Maximal Efficiency Transfection Results: Reporter Gene Plasmid Efficiency (GFP %) | collagen type I pre-treated culture dish ~90% DMEM with 4.5 g/L glucose, 10% FBS 3:1 Serum-free DMEM with 4.5 g/L glucose 15 minutes at RT Yes No 48 hours EGFP pEGFP-N3 (CMV promoter) 82% |
Storage ConditionStore at 4 °C. If stored properly, the product is stable for 12 months or longerA Picture Showing Transfection Efficiency of GenJet™ Reagent on HepG2 Cells
GenJet™ reagent is optimized for HepG2 cells (ATCC # HB-8065) with exceptional efficiency in comparison of Lipofectamine 2000 (L2K) and Amaxa electroporation device. HepG2 cells were grown per ATCC recommended culture medium on a collagen type I treated culture dish and transfected with pEGFP-N3 by GenJet™. The efficiency was checked 48 hours post transfection. Right Panel: Comparison of transfection efficiency of GenJet with Lipofecatmine 2000 (L2K), and Amaxa electroporation device on HepG2 cells. GFP DNA (pEGFP-N3, 4.7 kb) was introduced to HepG2 cell (cultured on Collagen pretreated dishes) with different transfection reagents per manufacturers protocols . GFP positive cell (%) and fluorescence intensity were detected by passing through FACS 48 hours post transfection Left Panel: presence of serum and antibiotics enhances GenJet reagents efficiency on HepG2 cells. HepG2 cell (grown on collagen treated dishes) was transfected with three different conditions-------serum and antibiotics free, presence of 10% serum and antibiotics followed by removal 5 hours post transfection and presence of 10% serum and antibiotics without removal 5 hours post transfection. GenJet™ reagent is optimized for HepG2 cells (ATCC # HB-8065). HepG2 cells were grown per ATCC recommended culture medium on a collagen type I treated culture dish and transfected with GFP (pEGFP-N3, 4.7 kb, right panel) and β-galactosidase cDNA (pSV-β-galactosidase, 6.9 kb, left panel) by pre-optimized GenJet™ DNA Transfection Reagent for HepG2 cells . The efficiency was checked 48 hours post transfection by Zeiss 510 Confocal Microscopy and β-galactosidase staining kit respectively. Data Sheet.pdf
