GenJet™ In Vitro DNA Tranfection Reagent for HepG2 Cells

Description

GenJet™ DNA In Vitro
Tranfection Reagent for HepG2 Cells is pre-optimized
for HepG2 cell transfection.

HepG2 (Hepatocellular carcinoma, human) is a perpetual add cell line which
was derived from the liver tissue of a 15 year old caucasian male with a well
differentiated hepatocellular carcinoma. These cells are epithelial in
morphology, have a model chromosome number of 55 and are not tumorigenic in nude
mice. The cells secrete a variety of major plasma proteins e.g.. albumin, alpha
2-macroglobulin, alpha 1-antitrypsin, transferrin and plasminogen. They
have been grown successfully in large scale cultivation systems. Hepatitis
B virus surface antigens have not been detected. HepG2 cells have been shown to
be G418 resistant (400µg/mL). The cells will respond to stimulation with human
growth hormone.

Refer to the following optimal transfection conditions for maximal transfection
efficiency on HepG2 cells. GenJet™ reagent, 1.0 ml, is sufficient for 300 to
600 transfections in 24 well plates or 150 to 300 transfections in 6 well plates.

Summary of Optimal Transfection Conditions:

Transfection Results:

Storage Condition
Store at 4 °C. If stored properly, the product is stable for 12 months or longer
A Picture Showing Transfection Efficiency of GenJet™ Reagent on HepG2 Cells

GenJet™ reagent is optimized for
HepG2 cells (ATCC # HB-8065)
with exceptional efficiency in comparison of Lipofectamine 2000 (L2K) and Amaxa
electroporation device.
HepG2 cells
were grown per ATCC recommended culture medium on a collagen type I treated
culture dish and transfected with pEGFP-N3 by
GenJet™. The efficiency was checked 48 hours post transfection.

Right Panel: Comparison of transfection efficiency of GenJet with Lipofecatmine
2000 (L2K),

and Amaxa electroporation
device on
HepG2 cells. GFP DNA (pEGFP-N3, 4.7 kb) was introduced to HepG2 cell (cultured
on Collagen pretreated dishes) with different transfection reagents per manufacturer’s protocols
. GFP positive cell (%)
and fluorescence intensity were detected by passing through FACS 48 hours post transfection

Left Panel: presence of serum and antibiotics enhances GenJet reagent’s
efficiency on HepG2 cells. HepG2 cell (grown on collagen treated dishes)
was transfected with three different conditions——-serum and antibiotics
free, presence of 10% serum and antibiotics followed by removal 5 hours post transfection and presence of 10% serum and antibiotics without removal 5 hours
post transfection.

GenJet™ reagent is optimized for
HepG2 cells (ATCC # HB-8065).
HepG2 cells
were grown per ATCC recommended culture medium on a collagen type I treated
culture dish and transfected with GFP (pEGFP-N3, 4.7 kb, right panel) and

β-galactosidase cDNA
(pSV-β-galactosidase,
6.9 kb, left panel) by pre-optimized GenJet™ DNA Transfection Reagent for HepG2
cells . The efficiency was checked 48 hours post transfection
by Zeiss 510 Confocal Microscopy and

β-galactosidase staining kit
respectively.

Data Sheet
To request a free trial sample, please
Create An Account with us to enter your shipping address and email us at order@signagen.com
Testimonials
It has been tested and better results were obtained than with Amaxa Nucleofactor.
A colleague and I, will be using it for all of our transfection experiments.
———-Fouad Francis., Ph.D., NICHD/NIH

I have got 70% efficiency with a GFP plasmid. With my protein of interest,
which is 200KDa, I got 20% efficiency, pretty comparable to Amaxa
electroporation. Thank you SignaGen.
——–Marina
Uhart, Ph.D., NHLBI/NIH
Wow!!!!! What a product. We have been using Lipofectamine 2000 on HepG2
and have been struggling to get even modest transfection efficiency. In
contrast, GenJet gave us dramatically higher transfection efficiency. By
eye, looks like 5-10 fold improvement. The especially impressive
part is that one uses less reagent and less DNA to get much better results. Our
future HepG2 experiments will now all be done with GenJet. Thanks.
——–Erik Snapp, Ph.D., Albert Einstein College of Medicine.