Description:
GenMute™ siRNA
Transfection Reagent for Insulinoma (INS) cell is pre-optimized for transfecting
siRNA to INS. Insulinoma cell is a tumor cell line derived from pancreatic beta
cells. Normal beta cells secrete insulin in response to increases in blood
glucose. The resulting increase in insulin acts to lower blood glucose back to
normal levels at which point further secretion of insulin is stopped. In
contrast, the secretion of insulin by INS cell is not properly regulated by
glucose and the tumors will continue to secrete insulin causing glucose levels
to fall further than normal.
Size & content:
- GenMute™ Reagent, 1.0 ml, sufficient for ~833 reactions based on transfecting
25 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
INS 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 (% ) |
RPMI 1640 with 4.5 g/L glucose, 10% FBS
~60%
50 nM
1xGenMute™
Transfection Buffer
~15 minutes at RT
48 hours
Renilla Luciferase
80% |
Storage:
Store at 4 °C. If stored properly, the product is stable for 12 months or
longer.
Data Sheet & Protocol