Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-N87 Transfection Reagent (Gastric Carcinoma, CRL5822) Altogen Biosystems offers the NCI-N87 Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The NCI-N87 Transfection Reagent is a two component formulation that enhances lipid mediated transfection efficiency, and it has been developed to provide high transfection efficiency with the NCI-N87 cell line. This cell line is a good host for a variety of research related to molecular and cellular biology. When cultured in vitro, NCI-N87 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-N87 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-N87 cells www.growth-media.net Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-N87 Transfection Reagent (Gastric Carcinoma, CRL5822) NCI-N87 Cell Line Description and Applications The NCI-N87 cell line originates from a metastatic site in the liver from a male patient suffering from gastric carcinoma. The cell line is unique due to its colony-forming growth pattern, expression of elevated levels of gastric zymogens and maintenance at post-confluency. The NCI-N87 cell line tests positive for erb B2 and myc oncogenes. The cell line also expresses receptors for muscarinic cholinergic agents. The NCI-N87 has applications for research related to stomach cancer because of its distinctive characteristics. NCI-N87 cells www.growth-media.net Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com

NCI-N87 Transfection Protocol Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-N87 Transfection Reagent (Gastric Carcinoma, CRL5822) NCI-N87 Transfection Protocol 1. Plate 10,000 - 15,000 NCI-N87 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-N87 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-N87 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-N87 Transfection Reagent (Gastric Carcinoma, CRL5822) 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-N87 Transfection Kit Product Details Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-N87 Transfection Reagent (Gastric Carcinoma, CRL5822) NCI-N87 Transfection Kit Product Details Two component formulation enhances lipid mediated transfection efficiency Optimized for intracellular delivery of plasmid DNA, siRNA, microRNA, and mRNA High transfection efficiency of both siRNA and plasmid DNA without compromising cell viability Achieve robust siRNA uptake for dependable gene silencing Effective transfection under conditions of up to 40% serum Transfection kit includes Transfection Enhancer reagent Gentle enough to be used for single cell analysis 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-N87 Transfection Reagent (Gastric Carcinoma, CRL5822) Data Figure 1. siRNAs targeting Lamin A/C mRNA or non-silencing control siRNA were transfected into NCI-N87 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).

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-N87 Transfection Reagent (Gastric Carcinoma, CRL5822) Data Figure 2. Protein expression of Lamin A in NCI-N87 cells. DNA plasmid expressing Lamin A or siRNA targeting Lamin A were transfected into NCI-N87 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-N87 Transfection Kit Benefits Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > NCI-N87 Transfection Reagent (Gastric Carcinoma, CRL5822) NCI-N87 Transfection Kit Benefits Pre-optimized transfection protocol for NCI-N87 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-N87 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-N87 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