Description
GenJet™ DNA In Vitro
Transfection Reagent for
Primary Macrophages is pre-optimized for transfecting 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. GenJet™ reagent, 1.0 ml, is
sufficient for 300 to 600 transfections in 24 well plates or 50 to 100
transfections in 6 well plates.
|
Summary
of Optimal Transfection Conditions:
Confluence on
the day of transfection
Cell culture
conditions
GenJet™ (µl) : DNA (µg) Ratio
Diluent for DNA and Transfection Reagent
Incubation Time to Form GenJet™/DNA Complex
Presence of Serum/Antibiotics during Transfection
Change Medium After Transfection
Maximal Efficiency
Transfection Results:
Reporter Gene
Plasmid
Efficiency (GFP %) |
~70%
PRMI with 10% FBS
3:1
1xGenJet™ Transfection Buffer
15 minutes at RT
Yes
No
48 hours
EGFP
pEGFP-N3 (CMV promoter)
75% |
Storage Condition
Store at 4 °C. If stored properly, the product is stable for 12 months or longer
Data Sheet & Protocol
