Description:
GenMute™
siRNA Transfection Reagent for U2OS Cell is pre-optimized for transfecting siRNA
to U2OS cells. The U2OS cell line, originally known as the 2T line, was cultivated from the
bone tissue of a fifteen-year-old human female suffering from osteosarcoma.
Established in 1964, the original cells were taken from a moderately
differentiated sarcoma of the tibia. U2OS cells exhibit epithelial
adherent morphology and viruses were not detected in the line during
co-cultivation with WI-38 cells or in CF tests against SV40, RSV, or
adenoviruses. Cells are positive for insulin-like growth factor I (IGF-I) and
insulin-like growth factor II (IGF II) receptors and express a number of
antigens, including blood type A, Rh+, HLA A2, Aw30, B12, Bw35, and B40(+/-).
Size & content:
-
GenMute™ Reagent,
1.0 ml, sufficient for ~833 reactions based on transfecting 15 pmoles
siRNA or miRNA mimics in 24-well plate
-
GenMute™ Transfection Buffer (5x ), formulated for maximal transfection efficiency, 8.0 ml
to make 40 ml working solution
Refer to the following optimal transfection conditions for maximal silencing on
U2OS cells.
GenMute™ reagent, 1.0 ml, is sufficient for ~833 transfections in 24 well plates or
~416 transfections in 6 well plates.
|
Summary of
Optimal Transfection Conditions:
Cell culture
medium
Confluency
of cell on the
day of transfection
Optimal siRNA concentration
Diluent for siRNA and GenMute™ Reagent
Incubation Time to Form GenMute™/siRNA
Complex
Maximal Silencing Efficiency
Transfection
Results:
Reporter Gene
Silencing
Efficiency (% ) |
DMEM with 4.5 g/L glucose, 10% FBS
~50%
30 nM
1xGenMute™
Transfection Buffer
~15 minutes at RT
48 hours
Renilla Luciferase
90% |
Storage:
Store at 4 °C. If stored properly, the product is stable for 12 months or
longer.
Data Sheet & Protocol
