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Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products > Neuro-2 a Transfection Reagent (Neuroblastoma Cells, CCL 131) Altogen Biosystems offers the Neuro-2 a Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The Neuro-2 a Transfection Reagent is a liposome formulation reagent, and it has been developed to provide high transfection efficiency with the Neuro-2 a cell line. This cell line is a good host for a wide variety of research in cell and molecular biology. When cultured in vitro, Neuro-2 a 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 Neuro-2 a 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 [email protected] com

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products > Neuro-2 a Transfection Reagent (Neuroblastoma Cells, CCL 131) Neuro-2 a Cell Line Description and Applications The Neuro-2 a cell line originates from brain tissue from an albino house mouse (Mus musculus) with neuroblastoma. The Neuro-2 a cell line displays amoeboid and neuronal stem cell morphology. The cell line produces large amounts of microtubular protein. Research indicates this protein is critical to a nerve cell contractile system that supports axoplasmic flow. The Neuro-2 a cell line is susceptible to several viruses: human poliovirus 1, vesicular stomatitis virus, and herpes simplex virus. This cell line makes an excellent transfection host, and its morphology has different applications in biological research. High Density Neuro-2 a Cells Atcc. com Altogen Biosystems 848 Rainbow Blvd #823 Las Vegas NV 89107 USA Telephone 702 349 6103 Fax 702 989 -0841 email [email protected] com

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products > Neuro-2 a Transfection Reagent (Neuroblastoma Cells, CCL 131) Neuro-2 a Transfection Protocol 1. Plate 10, 000 - 15, 000 Neuro-2 a cells per well in 0. 5 ml of complete growth medium 12– 24 hours prior to transfection 2. Wash with 1 x. PBS 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 m. RNA), or • 30 n. M - 50 n. M of si. RNA (or micro. RNA) *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 Neuro-2 a cells (from step 2) 7. Incubate cells at 37ºC in a humidified CO 2 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 Neuro-2 a 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 [email protected] com

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products > Neuro-2 a Transfection Reagent (Neuroblastoma Cells, CCL 131) 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 [email protected] com

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products > Neuro-2 a Transfection Reagent (Neuroblastoma Cells, CCL 131) Neuro-2 a Transfection Kit Product Details • Proprietary cationic lipids formulation • Transfection protocols provided for transfection of proteins, DNA, m. RNA, si. RNA, sh. RNA and micro. RNA • 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/si. RNA 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 [email protected] com

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products > Neuro-2 a Transfection Reagent (Neuroblastoma Cells, CCL 131) Data Figure 1. Cyclophilin B silencing efficiency was determined by q. RT-PCR in Neuro-2 a cells transfected by Cyclophilin B si. RNA or non-silencing si. RNA control following the recommended transfection protocol. Cyclophilin m. RNA expression levels were measured 48 hours post-transfection. 18 S r. RNA levels were used to normalize the Cyclophilin B data. Values are normalized to untreated sample. Data are presented as means ± SD (n=3).

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products > Neuro-2 a Transfection Reagent (Neuroblastoma Cells, CCL 131) Data Figure 2. Protein expression of Cyclophilin B in Neuro-2 a cells. DNA plasmid expressing Cyclophilin B or si. RNA targeting Cyclophilin B were transfected into Neuro-2 a 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

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products > Neuro-2 a Transfection Reagent (Neuroblastoma Cells, CCL 131) Neuro-2 a Transfection Kit Benefits • Pre-optimized transfection protocol for Neuro-2 a cell line • Compatible with DNA, small RNA (si. RNA, sh. RNA, mi. RNA), m. RNA, 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 Neuro-2 a 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 Neuro-2 a 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 [email protected] com