Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > SK-N-SH Transfection Reagent (Neuroblastoma Cells, HTB-11) Altogen Biosystems offers the SK-N-SH Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The SK-N-SH Transfection Reagent is an advanced cationic lipid formulation reagent, and it has been developed to provide high transfection efficiency with the SK-N-SH cell line. This cell line is a good host for studying brain cancer and cell and molecular biology. When cultured in vitro, SK-N-SH 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 SK-N-SH 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 SK-N-SH cells Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > SK-N-SH Transfection Reagent (Neuroblastoma Cells, HTB-11) SK-N-SH Cell Line Description and Applications The SK-N-SH cell line originates from a 4-year old female patient suffering from neuroblastoma. J. L. Biedler established this cell line from the brain tissue of the patient in 1970. The SK-N-SH cell line has a high population doubling time and also produces large amounts of dopamine-beta-hydroxylase. The cell has a modal chromosome number of 47, two x chromosomes (and therefore is genetically female), and also contains an abnormality: trisomy 1q. Research indicates this cell line is adenosinergic, glutamatergic and acetylcholinergic. After amyloid-beta peptide treatment, SK-N-SH will increase production of M-CSF. This cell is a good transfection host. High Density SK-N-SH 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

SK-N-SH Transfection Protocol Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > SK-N-SH Transfection Reagent (Neuroblastoma Cells, HTB-11) SK-N-SH Transfection Protocol 1. Plate 10,000 - 15,000 SK-N-SH 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 SK-N-SH 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 SK-N-SH 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 > SK-N-SH Transfection Reagent (Neuroblastoma Cells, HTB-11) 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

SK-N-SH Transfection Kit Product Details Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > SK-N-SH Transfection Reagent (Neuroblastoma Cells, HTB-11) SK-N-SH Transfection Kit Product Details A cationic lipid 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

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > SK-N-SH Transfection Reagent (Neuroblastoma Cells, HTB-11) Data Figure 1. siRNAs targeting Lamin A/C mRNA or non-silencing control siRNA were transfected into SK-N-SH 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 > SK-N-SH Transfection Reagent (Neuroblastoma Cells, HTB-11) Data Figure 2. Protein expression of Lamin A in SK-N-SH cells. DNA plasmid expressing Lamin A or siRNA targeting Lamin A were transfected into SK-N-SH 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.

SK-N-SH Transfection Kit Benefits Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > SK-N-SH Transfection Reagent (Neuroblastoma Cells, HTB-11) SK-N-SH Transfection Kit Benefits Pre-optimized transfection protocol for SK-N-SH 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 SK-N-SH 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 SK-N-SH 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