Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for 3LL Cells (Lewis Lung Carcinoma Cells) Altogen Biosystems offers the 3LL Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The 3LL Transfection Reagent is an advanced nanoparticle-based liposome formulation reagent, and it has been developed to provide high transfection efficiency with the 3LL cell line. This cell line is a good host for studying lung cancer and cell and molecular biology. When cultured in vitro, 3LL 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 3LL 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

3LL Cell Line Description and Applications Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for 3LL Cells (Lewis Lung Carcinoma Cells) 3LL or Lewis Lung Carcinoma cells are named after the cell biologist who discovered them, Dr. Margaret R. Lewis. She discovered this cell line at the Wistar Institute in 1951. The tumor arose from a spontaneous carcinoma from the lung of a C57BL mouse. 3LL cells are known for transplantable malignancy. This cell line can be used for studying lung cancer as well molecular and cell biology. Lewis Lung Carcinoma cells. Very recent research indicates that gene expression of 3LL metastatic cells allows for proliferation. Research indicates that the metastatic characteristic of this cell line can be diminished with ultraviolet treatment. Furthermore, studies show that cannabinoids can suppress 3LL tumor growth. 3LL Cell Line Description and Applications 3LL cells Lookfordiagnosis.com Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com

3LL Transfection Protocol Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for 3LL Cells (Lewis Lung Carcinoma Cells) 3LL Transfection Protocol 1. Plate 10,000 - 15,000 3LL 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 3LL 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 3LL 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

General Lipoplex-mediated Transfection Mechanism of Action Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for 3LL Cells (Lewis Lung Carcinoma Cells) 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

3LL Transfection Kit Product Details Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for 3LL Cells (Lewis Lung Carcinoma Cells) 3LL Transfection Kit Product Details A nanoparticle-based liposome 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

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for 3LL Cells (Lewis Lung Carcinoma Cells) Data Figure 1. GAPD mRNA levels were quantified using real-time RT-PCR in the cells transfected with siRNAs targeting GAPD or non-silencing siRNA. Forty-eight hours post-transfection, 3LL 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).

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for 3LL Cells (Lewis Lung Carcinoma Cells) Data Figure 2. Protein expression of GAPDH in 3LL cells. DNA plasmid expressing GAPDH or siRNA targeting GAPDH were transfected into 3LL 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.

3LL Transfection Kit Benefits Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Transfection Reagent for 3LL Cells (Lewis Lung Carcinoma Cells) 3LL Transfection Kit Benefits Pre-optimized transfection protocol for 3LL 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 3LL 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 3LL 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