Products > PC-12 Transfection Reagent (Rat Pheochromocytoma Cells) Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > PC-12 Transfection Reagent (Rat Pheochromocytoma Cells) Altogen Biosystems offers the PC-12 Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. PC-12 Transfection Reagent is a two component formulation that enhances lipid mediated transfection efficiency, and it is developed to provide high transfection efficiency with PC-12. This cell line is a good host for studying adrenal glands and related adrenal gland cancers in particular. Other applications include drug discovery, gene expression studies, molecular and cell biology research applications. Purchase PC-12 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 PC-12 Cell Line Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > PC-12 Transfection Reagent (Rat Pheochromocytoma Cells) PC-12 Cell Line Description and Applications The PC-12 cell line originates from a transplantable rat adrenal pheochromocytoma, and was established in 1976. The PC-12 cell line has a population doubling time is more than two days and will stop dividing and differentiate with a nerve growth factor treatment. In regards to differentiation, PC-12 cells will grow long neuritis and display cellular composition changes. The cells are small, have a rounded morphology, usually form in clusters and have limited cytoplasm. The PC-12 cell line is difficult to transfect, particularly after differentiation. This cell line has applications for studying adrenal glands and cancers related to adrenal glands. High Density PC-12 Cell Line 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

PC-12 Transfection Protocol Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > PC-12 Transfection Reagent (Rat Pheochromocytoma Cells) PC-12 Transfection Protocol Plate 10,000 - 15,000 PC-12 cells per well in 0.5 ml of complete growth medium 12–24 hours prior to transfection Wash with 1xPBS and add 0.5 ml of fresh growth medium 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 Incubate transfection complexes at RT for 15 - 30 minutes 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 Add prepared transfection complexes to 0.5 ml of complete growth medium with PC-12 cells (from step 2) Incubate cells at 37ºC in a humidified CO2 incubator 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 PC-12 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 > PC-12 Transfection Reagent (Rat Pheochromocytoma 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

PC-12 Transfection Kit Product Details Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > PC-12 Transfection Reagent (Rat Pheochromocytoma Cells) PC-12 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 > PC-12 Transfection Reagent (Rat Pheochromocytoma Cells) Data Figure 1. SiRNAs targeting Lamin A/C mRNA or non-silencing control siRNA were transfected into PC-12 cells following the recommended protocol. At 48 hours post-transfection the cells were analyzed by qRT-PCR for 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=3).

Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > PC-12 Transfection Reagent (Rat Pheochromocytoma Cells) Data Figure 2. Protein expression of Lamin A in PC-12 cells. DNA plasmid expressing Lamin A or siRNA targeting Lamin A were transfected into PC-12 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.

PC-12 Transfection Kit Benefits Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > PC-12 Transfection Reagent (Rat Pheochromocytoma Cells) PC-12 Transfection Kit Benefits Pre-optimized transfection protocol for PC-12 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 PC-12 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 PC-12 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