Description:
GenMute™
siRNA Transfection Reagent for PMG is pre-optimized for transfecting siRNA to
primary macrophages. Macrophages
are white blood cells within tissues, produced by the differentiation of monocytes. Monocytes and macrophages are phagocytes, acting
in both non-specific defense (innate immunity) as well as to help initiate
specific defense mechanisms (adaptive immunity) of vertebrate animals. Their
role is to phagocytose (engulf and then digest) cellular debris and pathogens
either as stationary or as mobile cells, and to stimulate lymphocytes and other
immune cells to respond to the pathogen. They can be identified by specific
expression of a number of proteins including CD14, CD11b, F4/80 (mice)/EMR1
(human), Lysozyme M, MAC-1/MAC-3 and CD68 by flow cytometry or
immunohistochemical staining.
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
primary macrophages. GenMute™ reagent, 1.0 ml, is sufficient for ~833 transfections in 24 well plates or
~416 transfections in 6 well plates.
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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
~90%
50 nM
1xGenMute™
Transfection Buffer
~15 minutes at RT
48 hours
Renilla Luciferase
85% |
Storage:
Store at 4 °C. If stored properly, the product is stable for 12 months or
longer.
Data Sheet & Protocol
