Contents Per Kit:
●
1x1.0 ml of GenJet™
DNA Transfection Reagent
● 1x8.0 ml (x5 ) of
HapiCell™ Transfection
Toxicity Removal Cocktail
Description
GenJet™ DNA In Vitro
Transfection Reagent for SaoS-2 Cells is pre-optimized for transfecting human
epithelial-like osteosarcoma cell line, SaoS-2. SaoS-2 (sarcoma osteogenic)
are a non-transformed cell line derived from the primary osteosarcoma of an
11-year old Caucasian girl (since identified as Alice) in 1973 by Fogh et al[1].
In 1987, Rodan determined that Saos-2 cells "possess several osteoblastic
features and could be useful as a permanent line of human osteoblast-like cells
and as a source of bone-related molecules". With our pripariotary
transfection booster conjugation technology, this reagent was formulated to transfect
Saos-2 cells
with up to 60% efficiency (GFP positive cells).
SaoS-2 cell is one of
hard-to-transfect cell lines partly due to the transfection
toxicity related cell death. In combination
of our proprietary HapiCell™ Toxicity Removal Cocktail, the pre-optimized transfection
reagent gave up to 60% efficiency.
Refer to the following optimal transfection conditions for maximal transfection
efficiency on SaoS-2 cells.
To achieve maximal efficiency,
the "Shaved Cell
Transfection" protocol was optimized for transfecting SaoS-2 cells. Refer to the following optimal transfection conditions for maximal transfection
efficiency on SaoS-2 cells. GenJet™ reagent, 1.0 ml, is sufficient for 300 transfections in 24 well plates or 150 transfections in 6 well plates.
|
Summary of
Optimal Shaved CellTransfection Conditions:
Cell culture
medium
Confluency
of cell on the
day of shaving
Cell density for plating
GenJet™ (µl) : DNA (µg) Ratio
Diluent for DNA and Transfection Reagent
Incubation Time to Form GenJet™/DNA
Complex
Incubation time of shaved cells with transfection complex
Maximal Efficiency
Transfection
Results:
Reporter Gene
Plasmid
Efficiency (GFP %) |
DMEM with 4.5 g/L glucose, 10% FBS
95~100%
1.3x10^5 cells per square centimeter
4:1
Serum-free DMEM with 4.5 g/L glucose
15 minutes at RT
20 minutes at 37 degree
48 hours
EGFP
pEGFP-N3 (CMV promoter)
60% |
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 SaoS-2 Cells.
Saos-2 cells
were grown per ATCC recommended culture medium and transfected with GFP cDNA (pEGFP-N3) by GenJet™. The efficiency was checked 24 hours post transfection
by Nikon Elipse 2000 confocal microscopy
Data Sheet
To request a free trial sample, please
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us at
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