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.
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