Products > NCI-H441 Transfection Reagent (Lung Adenocarcinoma) Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-H441 Transfection Reagent (Lung Adenocarcinoma) Altogen Biosystems offers the NCI-H441 Cell Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The NCI-H441 Cell Transfection Reagent is a biodegradable polymer-based transfection reagent, and it has been developed to provide high transfection efficiency with NCI-H441. This cell line is a good host for studying a range of areas within the field of biomedical therapeutics and biology as well as lung cancer. When cultured in vitro, NCI-H441 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-H441 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

NCI-H441 Cell Line Description and Applications Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-H441 Transfection Reagent (Lung Adenocarcinoma) NCI-H441 Cell Line Description and Applications D. Carney, M. Brower, and A.F. Gazdar established the NCI-H441 cell line in 1982. The cell line was derived from pericardial fluid from a male patient with a lung papillary adenocarcinoma. The NCI-H441 cell line displays epithelial morphology and expresses mRNA, major surfactant apoprotein as well as pulmonary surfactant protein. This cell line can be cloned in soft agar with serum or without serum. Research with electron microscopy indicates that the cell line has multilamellar bodies and structures similar to Clara cell granules. NCI-H441 cells Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com

NCI-H441 Transfection Protocol Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-H441 Transfection Reagent (Lung Adenocarcinoma) NCI-H441 Transfection Protocol 1. Plate 10,000 - 15,000 NCI-H441 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-H441 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-H441 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 > NCI-H441 Transfection Reagent (Lung Adenocarcinoma) 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

NCI-H441 Transfection Kit Product Details Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-H441 Transfection Reagent (Lung Adenocarcinoma) NCI-H441 Transfection Kit Product Details A biodegradable polymer-based transfection reagent – once inside the cell, the polymer degrades into smaller less toxic components reducing cell toxicity, facilitating release of the transgene, and improving transfection efficiency Optimized easy-to-use transfection protocol provided for transfection of siRNA, DNA, mRNA, and microRNA Kit includes Transfection Enhancer reagent and recommended transfection protocol High transfection efficacy in the presence of serum Expand your RNAi application with a reagent optimized for delivery of both siRNA and plasmid Reproducible transfection results Works well for standard reverse transfection and high-throughput applications 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

Data Products > NCI-H441 Transfection Reagent (Lung Adenocarcinoma) Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-H441 Transfection Reagent (Lung Adenocarcinoma) Data Figure 1. siRNAs targeting Lamin A/C mRNA or non-silencing control siRNA were transfected into NCI-H441 cells following the recommended protocol. At 48 hours post-transfection the cells were analyzed by qRT-PCR for Lamin A/C gene expression levels. 18S rRNA levels were used to normalize the Lamin A/C data. Values are normalized to untreated sample. Data are means ± SD (n=4).

Data Products > NCI-H441 Transfection Reagent (Lung Adenocarcinoma) Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-H441 Transfection Reagent (Lung Adenocarcinoma) Data Figure 2. Protein expression of Lamin A in NCI-H441 cells. DNA plasmid expressing Lamin A or siRNA targeting Lamin A were transfected into NCI-H441 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.

NCI-H441 Transfection Kit Benefits Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-H441 Transfection Reagent (Lung Adenocarcinoma) NCI-H441 Transfection Kit Benefits Pre-optimized transfection protocol for NCI-H441 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-H441 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-H441 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