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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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HUH-7 Transfection Reagent (Liver Cancer Cells)
Altogen Biosystems offers the HUH-7 Transfection Reagent
among a host of 100+ cell line specific In Vitro Transfection Kits.
The HUH-7 Transfection Reagent is an advanced biodegradable
polymer based transfection reagent that enhances lipid mediated
transfection, and it has been developed to provide high
transfection efficiency with HUH-7 cells.
This cell line is a good host for studying liver cancer. When
cultured in vitro, HUH-7 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 HUH-7 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
Products >
HUH-7 Transfection Reagent (Liver Cancer Cells)
The HUH-7 cell line originates from the liver tumor of a
57-year old male Japanese patient. The cell line was
established in 1982 by Nakabayshi, H. and Sato, J. The
HUH-7 cell line is a well differentiated cellular carcinoma
cell line that is a type of immortal cell line. As the cell line
is a hepatoma, the cells display epithelial cell morphology.
Liver cancer can occur commonly as a result of cirrhosis
(alcohol-induced) or viral hepatitis infection.
HUH-7 Cells
cellbank.nibiohn.go.jp
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
HUH-7 Transfection Reagent (Liver Cancer Cells)
HUH-7 Transfection Protocol
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1. Plate 10,000 - 15,000 HUH-7 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 HUH-7 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 HUH-7 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
HUH-7 Transfection Reagent (Liver Cancer Cells)
General Lipoplex-mediated Transfection Mechanism of Action
Products >
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
HUH-7 Transfection Reagent (Liver Cancer Cells)
HUH-7 Transfection Kit Product Details
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•
A biodegradable polymer based transfection reagent – once inside the cell, the polymer degrades
into smaller less toxic components reducing cell toxicity, facilitating release of the transgene, and
improving transfection efficiency
•
Transfection protocols provided for transfection of proteins, DNA, mRNA, siRNA, shRNA and
microRNA
•
Transfection Enhancer reagent provided with the kit
•
Produce higher level of recombinant protein expression with minimal disruption of normal cell
function
•
Generate physiologically relevant data you can trust
•
Effective for plasmid DNA/siRNA co-transfection
•
Easy-to-use transfection protocol with reproducible results
•
Low cytotoxicity
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 >
HUH-7 Transfection Reagent (Liver Cancer Cells)
Data
Figure 1. GAPD mRNA levels were quantified using real-time RT-PCR in the Huh-7 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 >
HUH-7 Transfection Reagent (Liver Cancer Cells)
Data
Figure 2. Protein expression of GAPDH in HUH-7 cells. DNA plasmid expressing GAPDH or siRNA targeting
GAPDH were transfected into HUH-7 cells following Altogen Biosystems transfection protocol. At 72 hours posttransfection 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
HUH-7 Transfection Reagent (Liver Cancer Cells)
HUH-7 Transfection Kit Benefits
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• Pre-optimized transfection protocol for HUH-7 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 HUH-7 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 HUH-7 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]