Products > NCI-H1299 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-H1299 Transfection Reagent (Lung Adenocarcinoma) Altogen Biosystems offers the NCI-H1299 Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. NCI-H1299 Transfection Reagent is an advanced formulation of lipid based reagent, and it has been developed to provide high transfection efficiency with NCI-H1299 and other lung cancer cell lines. This cell line is a good host for studying lung cancer and related areas of study. When cultured in vitro, NCI-H1299 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-H1299 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

High Density NCI-H1299 Cells atcc.org Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-H1299 Transfection Reagent (Lung Adenocarcinoma) NCI-H1299 Cell Line Description and Applications The NCI-H1299 line originates from the lung cells of a 43-year old Caucasian male patient suffering from non-small cell lung cancer. The patient received radiation prior to cell line establishment. The cell line is specifically derived from a lymph node metastatic site. The NCI-H1299 cell line displays epithelial morphology and stain positive for keratin and vimentin and produce neuromedin B. The NCI-H1299 cell line has a partial deletion of the p53 protein and lacks expression of the p53 protein. This cell line is particularly useful research related to lung cancer. High Density NCI-H1299 Cells atcc.org Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com

NCI-H1299 Transfection Protocol Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-H1299 Transfection Reagent (Lung Adenocarcinoma) NCI-H1299 Transfection Protocol 1. Plate 10,000 - 15,000 NCI-H1299 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-H1299 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-H1299 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-H1299 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-H1299 Transfection Kit Product Details Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-H1299 Transfection Reagent (Lung Adenocarcinoma) NCI-H1299 Transfection Kit Product Details Two component formulation enhances lipid mediated 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

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-H1299 Transfection Reagent (Lung Adenocarcinoma) Data Figure 1. GAPD mRNA levels were quantified using real-time RT-PCR in the NCI-H1299 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 GAPD 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=5).

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-H1299 Transfection Reagent (Lung Adenocarcinoma) Data Figure 2. Protein expression of GAPDH in NCI-H1299 cells. DNA plasmid expressing GAPDH or siRNA targeting GAPDH were transfected into NCI-H1299 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-H1299 Transfection Kit Benefits Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-H1299 Transfection Reagent (Lung Adenocarcinoma) NCI-H1299 Transfection Kit Benefits Pre-optimized transfection protocol for NCI-H1299 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-H1299 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-H1299 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