HepG2 Transfection Reagent - High-efficiency transfection for Hepatocellular Carcinoma cells

1. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products >HepG2 Transfection Reagent (Hepatocellular Carcinoma) Altogen Biosystems offers the HepG2 Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The HepG2 Transfection Reagent is a proprietary cationic lipids formulation that enhances lipid-mediated transfection, and it has been developed to provide high transfection efficiency with HepG2 cells. This cell line is a good host for transfection and research related to liver function. When cultured in vitro, HepG2 cells are a monolayer and adhere to the surface of the culture flask. Other applications include drug discovery, gene expression studies, molecular and cell biology research applications. Purchase HepG2 Transfection Kit at www.Altogen.com Altogen Biosystems 848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone 702 349 6103  Fax 702 989-0841  email  techserv@altogen.com

2. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products >HepG2 Transfection Reagent (Hepatocellular Carcinoma) HepG2 Cell Line Description and Applications The HepG2 cell line originates from liver tissue of a 15-year-old male patient who suffered from differentiated hepatocellular carcinoma. The HepG2 cell line and derivatives can be used as a model system for research related to liver metabolism as well as hepatocarcinogenesis, xenobiotics toxicity, antigenotoxic and/or cogenotoxic agents. These cells display epithelial morphology and have a model chromosome number of 55. HepG2 cells produce albumin, plasminogen, fibrinogen, alpha 2-macroglobulin, alpha 1-antitrypsin, and transferrin. Research indicates that HepG2 cells are resistant to G418, but are responsive to human growth hormone stimulation. HepG2 Cells Altogen.com Altogen Biosystems 848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone 702 349 6103  Fax 702 989-0841  email  techserv@altogen.com

3. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products >HepG2 Transfection Reagent (Hepatocellular Carcinoma) HepG2 Transfection Protocol 1. Plate 10,000 - 15,000 HepG2 cells per well in 0.5 ml of complete growth medium 12–24 hours prior to transfection 2. Wash with 1xPBS and add 0.5 ml of fresh growth medium 3. Prepare transfection complexes by mixing 40 µl of serum-free medium, 5.5 µl of transfection reagent, and • 750 ng DNA (or mRNA), or • 30 nM - 50 nM of siRNA (or microRNA) *Referred to a final volume including growth medium 4. Incubate transfection complexes at RT for 15 - 30 minutes 5. Optional: Add 2 µl of Complex Condenser. This reagent reduces the size of transfection complex, therefore increasing transfection efficiency; however it may increase cell toxicity 6. Add prepared transfection complexes to 0.5 ml of complete growth medium with HepG2 cells (from step 2) 7. Incubate cells at 37ºC in a humidified CO2 incubator 8. Assay for phenotype or target gene expression 48 - 72 hours after transfection Optional: Transfection efficiency can be increased by addition of Transfection Enhancer reagent. Add 2 µl of Transfection Enhancer reagent 12-24 hours after transfection If the viability of HepG2 cells being transfected is affected at 16 - 24 hours post-transfection, the level of cytotoxicity can be decreased by changing the growth medium and eliminating redundant exposure of cells to transfectant Altogen Biosystems 848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone 702 349 6103  Fax 702 989-0841  email  techserv@altogen.com

4. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products >HepG2 Transfection Reagent (Hepatocellular Carcinoma) General Lipoplex-mediated Transfection Mechanism of Action Altogen Biosystems 848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone 702 349 6103  Fax 702 989-0841  email  techserv@altogen.com

5. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products >HepG2 Transfection Reagent (Hepatocellular Carcinoma) HepG2 Transfection Kit Product Details • Proprietary cationic lipids formulation • Transfection protocols provided for transfection of proteins, DNA, mRNA, siRNA, shRNA and microRNA • Transfection Enhancer reagent provided with the kit • Produce higher level of recombinant protein expression with minimal disruption of normal cell function • Generate physiologically relevant data you can trust • Effective for plasmid DNA/siRNA co-transfection • Easy-to-use transfection protocol with reproducible results • Low cytotoxicity • Developed and manufactured by Altogen Biosystems (www.Altogen.com) Altogen Biosystems 848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone 702 349 6103  Fax 702 989-0841  email  techserv@altogen.com

6. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products >HepG2 Transfection Reagent (Hepatocellular Carcinoma) Data Figure 1. GAPD mRNA levels were quantified using real-time RT-PCR in the HepG2 cells transfected with siRNAs targeting GAPD or non-silencing siRNA. Forty-eight hours post-transfection, the cells were harvested and analyzed by real-time RT-PCR for GAPDH mRNA expression levels. Data were normalized against the 18S rRNA signal. Control samples were either mock-transfected or untreated. Values are normalized to untreated sample. Data are means ± SD (n=3).

7. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products >HepG2 Transfection Reagent (Hepatocellular Carcinoma) Data Figure 2. Protein expression of GAPDH in HepG2 cells. DNA plasmid expressing GAPDH or siRNA targeting GAPDH were transfected into HepG2 cells following Altogen Biosystems transfection protocol. At 72 hours post-transfection the cells were analyzed by Western Blot for protein expression levels (normalized by total protein, 10 µg of total protein loaded per each well). Untreated cells used as a negative control.

8. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products >HepG2 Transfection Reagent (Hepatocellular Carcinoma) HepG2 Transfection Kit Benefits • Pre-optimized transfection protocol for HepG2 cell line • Compatible with DNA, small RNA (siRNA, shRNA, miRNA), mRNA, and small protein complexing • Free of serum and protein of animal origin • Compatible with standard and reverse transfection methods (both protocols provided in the kit manual) • Easy to use HepG2 transfection protocol ensures great performance with expedited experimental timeline • Equally efficient for single or multiple transfections • Can be used for transient transfection and development of stable HepG2 cell lines • Bio-degradable after endocytosis • Used for preclinical research worldwide Altogen Biosystems 848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone 702 349 6103  Fax 702 989-0841  email  techserv@altogen.com