GenJet™ DNA In Vitro Tranfection Reagent for K562 is pre-optimized for transfecting K562 cells. K562 cells were the first human immortalised myelogenous leukaemia line to be established and are a bcr:abl positive erythroleukamia line derived from a 53 year old female CML patient in blast crisis. The cells are non-adherent and rounded, and bear some proteomic resemblance to both undifferentiated granulocytes and erythrocytes. In culture they exhibit much less clumping than many other suspension lines, presumably due to the downregulation of surface adhesion molecules by bcr:abl. K562s can spontaneously develop characteristics similar to early-stage erythrocytes, granulocytes and monocytes and are easily killed by natural killer cells as they lack the MHC complex required to inhibit NK activity. They also lack any trace of Epstein-Barr virus and other herpesviruses. In addition to the Philadelphia chromosome they also exhibit a second reciprocal translocation between the long arm of chromosome 15 with chromosome 17.

Refer to the following optimal transfection conditions for maximal transfection efficiency on K562 cells. 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
Efficiency (GFP %)

DMEM with 4.5 g/L glucose, 10% FBS

Serum-free DMEM with 4.5 g/L glucose

15 minutes at RT
Yes, ~5 Hours After Transfection
48 hours

pEGFP-N3 (CMV promoter) 


Storage Condition
Store at 4 °C. If stored properly, the product is stable for 12 months or longer

Data Sheet 

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