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Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
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NIH3T3 Transfection Reagent (Swiss Mouse Fibroblasts)
Altogen Biosystems offers the NIH3T3 Transfection Reagent
among a host of 100+ cell line specific In Vitro Transfection Kits.
NIH3T3 Transfection Reagent is an Proprietary cationic lipids
formulation, and it is developed to provide high transfection
efficiency with NIH3T3.
This cell line is a good host for studying mouse viruses and human
embryonic stem cells. When cultured in vitro, NIH3T3 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 NIH3T3 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
NIH3T3 Transfection Reagent (Swiss Mouse Fibroblasts)
NIH3T3 Cell Line Description and Applications
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Image of NIH3T3 Cells
Altogen.com
The NIH3T3 cell line was established in 1962 by
researchers at the New York University School of
Medicine. It was derived from cells taken from a Swiss
mouse embryo. The 3T3 cell line is a type of murine
fibroblast cell and is now the classic fibroblast cell line in
research. Furthermore, the cell line can be used as “feeder
cells” for research related and maintenance of
undifferentiated human embryonic stem cells. Murine
embryonic fibroblasts, and in particular the 3T3 cell line
can be used to secrete growth factors to cultivate
keratinocytes. Research indicates that NIH3T3 cells are
susceptible to the leukemia and sarcoma viruses and that
benzodiazepines inhibit growth of this cell line. The 3T3
cell line is an excellent transfection host and is also useful
for titration studies related to mouse viruses.
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
NIH3T3 Transfection Reagent (Swiss Mouse Fibroblasts)
NIH3T3 Transfection Protocol
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1. Plate 10,000 - 15,000 NIH3T3 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 NIH3T3 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 NIH3T3 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
NIH3T3 Transfection Reagent (Swiss Mouse Fibroblasts)
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
NIH3T3 Transfection Reagent (Swiss Mouse Fibroblasts)
NIH3T3 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 >
NIH3T3 Transfection Reagent (Swiss Mouse Fibroblasts)
Data
Figure 1. GAPD mRNA levels were quantified using real-time RT-PCR in NIH3T3 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 GAPDH 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=3).
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products >
NIH3T3 Transfection Reagent (Swiss Mouse Fibroblasts)
Data
Figure 2. Protein expression of GAPDH in NIH3T3 cells. DNA plasmid expressing GAPDH or siRNA targeting
GAPDH were transfected into NIH3T3 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
NIH3T3 Transfection Reagent (Swiss Mouse Fibroblasts)
NIH3T3 Transfection Kit Benefits
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• Pre-optimized transfection protocol for NIH3T3 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 NIH3T3 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 NIH3T3 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]