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Transfection with competitor reagent was performed according to manufacturer’s instructions
BHK21 cells were seeded from 90-100% confluent culture the day before transfection with the density 10,000 cells per well in 100 μl complete growth medium DMEM + 10% Fetal Bovine Serum (Gibco®).
Optimal ratio of reagent/DNA varies from 0.25μl/0.1μg to 0.4μl/0.1μg.
SEAP activity is quantitatively determined by a colorimetric method based on hydrolysis
of chromogenic substrate para-nitrophenyl phosphate (PNPP). 1 μg/μl of PNPP is
prepared in 1M Diethanolamine, 1mM MgCl2, pH9.8.