Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Detroit-551 Transfection Reagent (Embryonic Fibroblast Cells) Altogen Biosystems offers the Detroit-551 Cell Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The Detroit-551 Cell Transfection Reagent is a biodegradable polymer-based transfection agent, and it has been developed to provide high transfection efficiency with Detroit-551. This cell line is a good host for studying a range of areas within the field of biomedical therapeutics and biology. When cultured in vitro, Detroit-551 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 Detroit-551 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

Detroit-551 Cells ezbiosystems.com Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Detroit-551 Transfection Reagent (Embryonic Fibroblast Cells) Detroit-551 Cell Line Description and Applications The Detroit-551 cell line originates from the skin tissue of a generally healthy fetus from a Caucasian female taken in 1965. The cell line displays fibroblast morphology and cell type. The cell line has a lifespan of approximately 25 serial passages when taken from the tissue of origin. The Detroit-551 cell line growth can, however, be enhanced with a tumor necrosis factor alpha medium, making it an essential tool in research. The cell line can be transformed by SV40. It is also susceptible to Human poliovirus 1, Herpes simplex virus, and Vesicular stomatitis virus. Detroit-551 Cells ezbiosystems.com Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA Telephone  702 349 6103  Fax  702 989-0841  email  techserv@altogen.com

Detroit-551 Transfection Protocol Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Detroit-551 Transfection Reagent (Embryonic Fibroblast Cells) Detroit-551 Transfection Protocol 1. Plate 10,000 - 15,000 Detroit-551 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 Detroit-551 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 Detroit-551 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 > Detroit-551 Transfection Reagent (Embryonic Fibroblast Cells) 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

Detroit-551 Transfection Kit Product Details Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Detroit-551 Transfection Reagent (Embryonic Fibroblast Cells) Detroit-551 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 > Detroit-551 Transfection Reagent (Embryonic Fibroblast Cells) Data Figure 1. siRNAs targeting Lamin A/C mRNA or non-silencing control siRNA were transfected into Detroit-551 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 > Detroit-551 Transfection Reagent (Embryonic Fibroblast Cells) Data Figure 2. Protein expression of Lamin A/C in Detroit-551 cells. DNA plasmid expressing Lamin A/C or siRNA targeting Lamin A/C were transfected into Detroit-551 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.

Detroit-551 Transfection Kit Benefits Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > Detroit-551 Transfection Reagent (Embryonic Fibroblast Cells) Detroit-551 Transfection Kit Benefits Pre-optimized transfection protocol for Detroit-551 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 Detroit-551 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 Detroit-551 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