Transcript PPT - Altogen Biosystems

Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
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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 a good 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  [email protected]
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
C2C12 Transfection Reagent (Mouse Myoblast Cells)
C2C12 Cell Line Description and Applications
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High Density C2C12 Cell Line
atcc.org
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).
Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA
Telephone  702 349 6103  Fax  702 989-0841  email  [email protected]
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
C2C12 Transfection Reagent (Mouse Myoblast Cells)
C2C12 Transfection Protocol
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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  [email protected]
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
C2C12 Transfection Reagent (Mouse Myoblast Cells)
General Lipoplex-mediated Transfection Mechanism of Action
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Altogen Biosystems  848 Rainbow Blvd #823  Las Vegas  NV  89107  USA
Telephone  702 349 6103  Fax  702 989-0841  email  [email protected]
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
C2C12 Transfection Reagent (Mouse Myoblast Cells)
C2C12 Transfection Kit Product Details
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•
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  [email protected]
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).
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.
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
C2C12 Transfection Reagent (Mouse Myoblast Cells)
C2C12 Transfection Kit Benefits
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• 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  [email protected]