NCI-H460 Transfection Reagent - High Efficiency Lung Carcinoma Transfection Kit

1. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-H460 Transfection Reagent (Lung Carcinoma, HTB177) Altogen Biosystems offers the NCI-H460 Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The NCI-H460 Transfection Reagent is a proprietary cationic lipids formulation, and it has been developed to provide high transfection efficiency with the NCI-H460 cell line. This cell line is a good host for studying lung cancer. When cultured in vitro, NCI-H460 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 NCI-H460 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 > NCI-H460 Transfection Reagent (Lung Carcinoma, HTB177) NCI-H460 Cell Line Description and Applications A. F. Gazdar and associates established the NCI-H460 cell line in 1982 from tissue derived from the pleural fluid of a male patient with large cell lung carcinoma. This cell line is tumorigenic and displays epithelial morphology. This cell line is unique because it does not appear to have significant DNA abnormalities. As normal long tissue does, the NCI-H460 cell line can express easily detectable p53 mRNA. The NCI-H460 cell line is negative for neurofilament triplet protein and stains positively for vimentin and keratin. This cell line has applications for biomedical research related to lung cancer. NCI-H460 cells nature.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 > NCI-H460 Transfection Reagent (Lung Carcinoma, HTB177) NCI-H460 Transfection Protocol 1. Plate 10,000 - 15,000 NCI-H460 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 NCI-H460 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 NCI-H460 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 > NCI-H460 Transfection Reagent (Lung Carcinoma, HTB177) 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 > NCI-H460 Transfection Reagent (Lung Carcinoma, HTB177) NCI-H460 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 > NCI-H460 Transfection Reagent (Lung Carcinoma, HTB177) Data Figure 1. Cyclophilin B silencing efficiency was determined by qRT-PCR in the NCI-460 cells transfected by Cyclophilin B siRNA or non-silencing siRNA control following the recommended transfection protocol. Cyclophilin mRNA expression levels were measured 48 hours post-transfection. 18S rRNA levels were used to normalize the Cyclophilin B data. Values are normalized to untreated sample. Data are presented as means ± SD (n=6).

7. Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-H460 Transfection Reagent (Lung Carcinoma, HTB177) Data Figure 2. Protein expression Cyclophilin B in NCI-H460 cells. DNA plasmid expressing Cyclophilin B or siRNA targeting Cyclophilin B were transfected into NCI-H460 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 > NCI-H460 Transfection Reagent (Lung Carcinoma, HTB177) NCI-H460 Transfection Kit Benefits • Pre-optimized transfection protocol for NCI-H460 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 NCI-H460 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 NCI-H460 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