Products > C2C12 Transfection Reagent (Mouse Myoblast Cells) Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > C2C12 Transfection Reagent (Mouse Myoblast Cells) Altogen Biosystems offers the C2C12 Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. C2C12 Transfection Reagent is an advanced formulation of a lipid-based reagent, and it has been developed to provide high transfection efficiency with C2C12 and other mammalian myoblast cells. This cell line is an excellent host for biomedical research. When cultured in vitro, C2C12 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 C2C12 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 C2C12 Cell Line atcc.org Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > C2C12 Transfection Reagent (Mouse Myoblast Cells) C2C12 Cell Line Description and Applications The C2C12 cell line was established by H. Blau, et al. as a subclone of the mouse (Mus musculus) myoblast cell line derived by D. Yaffe and O. Saxel. The C2C12 cell line was taken from muscle tissue and displays myoblast morphology. Although this cell line is an excellent transfection host, it differentiates rapidly and can form contractile myotubes that produce proteins associated with muscle. Furthermore, treating C2C12 cells with bone morphogenic protein 2 can shift this differentiation pathway to osteoblastic rather than myoblastic. The C2C12 cell line tests negative for the mouse pox virus (ectromelia virus). High Density C2C12 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

C2C12 Transfection Protocol Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > C2C12 Transfection Reagent (Mouse Myoblast Cells) C2C12 Transfection Protocol 1. Plate 10,000 - 15,000 C2C12 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 C2C12 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 C2C12 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 > C2C12 Transfection Reagent (Mouse Myoblast 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

C2C12 Transfection Kit Product Details Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > C2C12 Transfection Reagent (Mouse Myoblast Cells) C2C12 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

Data Products > C2C12 Transfection Reagent (Mouse Myoblast Cells) Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > C2C12 Transfection Reagent (Mouse Myoblast Cells) Data Figure 1. GAPD mRNA levels were quantified using real-time RT-PCR in the C2C12 cells transfected with siRNAs targeting GAPD or non-silencing siRNA. Forty-eight hours post-transfection, the cells were harvested and analyzed by real-time RT-PCR for GAPD mRNA expression levels. Data were normalized against the 18S rRNA signal. Control samples were either mock-transfected or untreated. Values are normalized to untreated sample. Data are means ± SD (n=5).

Data Products > C2C12 Transfection Reagent (Mouse Myoblast Cells) Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > C2C12 Transfection Reagent (Mouse Myoblast Cells) Data Figure 2. Protein expression of GAPDH in C2C12 cells. DNA plasmid expressing GAPDH or siRNA targeting GAPDH were transfected into C2C12 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.

C2C12 Transfection Kit Benefits Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products > C2C12 Transfection Reagent (Mouse Myoblast Cells) C2C12 Transfection Kit Benefits Pre-optimized transfection protocol for C2C12 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 C2C12 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 C2C12 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