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Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > Hep-3B
Transfection Reagent (Hepatocellular Carcinoma)
Altogen Biosystems offers the Hep-3B Transfection Reagent
among a host of 100+ cell line specific In Vitro Transfection
Kits.
The Hep-3B Transfection Reagent is a nanoparticle-based
liposome formulation, and it has been developed to provide
high transfection efficiency with the Hep-3B cell line.
This cell line is a good host for studying therapeutics for
Hepatitis B as well as cancers. When cultured in vitro, Hep3B 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 Hep-3B 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 > Hep-3B
Transfection Reagent (Hepatocellular Carcinoma)
Hep-3B Cell Line Description and Applications
Hep-3B cells
Youtube.com
The Hep-3B cell line originates from the
liver tissue of a black 8-year old suffering
from hepatocellular carcinoma. This cell line
displays epithelial morphology and makes
an excellent transfection host for in vitro
models related to liver biology. The Hep-3B
cell line is particularly useful for research
related to cancer as well as therapeutics for
hepatitis B because the Hep-3B cell line
genome contains the Hepatitis B virus. This
cell line has a modal number of 60 and has
a rearranged chromosome 1.
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 > Hep-3B
Transfection Reagent (Hepatocellular Carcinoma)
Hep-3B Transfection Protocol
1. Plate 10,000 - 15,000 Hep-3B 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 Hep-3B 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 Hep-3B 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
Products > Hep-3B
Transfection Reagent (Hepatocellular Carcinoma)
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  [email protected]
Leading Developer and Manufacturer of In Vivo and DNA Transfection
Kits, Transfection Reagents and Electroporation Delivery Products
Products > Hep-3B
Transfection Reagent (Hepatocellular Carcinoma)
Hep-3B Transfection Kit Product Details
•A nanoparticle-based liposome formulation
•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 > Hep-3B
Transfection Reagent (Hepatocellular Carcinoma)
Data
Figure 1. GAPD mRNA levels were quantified using real-time RT-PCR in the Hep-3B 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 > Hep-3B
Transfection Reagent (Hepatocellular Carcinoma)
Data
Figure 2. Protein expression of GAPDH in Hep-3B cells. DNA plasmid expressing GAPDH or siRNA targeting
GAPDH were transfected into Hep-3B 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
Products > Hep-3B
Transfection Reagent (Hepatocellular Carcinoma)
Hep-3B Transfection Kit Benefits
• Pre-optimized transfection protocol for Hep-3B 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 Hep-3B 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 Hep-3B 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]