Criterion precast sds polyacrylamide gels

We prepared whole tissue lysates of prostate, liver, lungs and spleen of PKCε^LoxP/LoxP and PKCε^LoxP/LoxP/PB^Cre4/+ mice as described previously [40 (link)]. Fifty micrograms of cell lysate were fractionated on 10-15% Criterion precast SDS-polyacrylamide gels (Bio-Rad Laboratories, Hercules CA). The fractionated proteins were transferred to 0.45 μm Hybond-P polyvinylidene difluoride (PVDF) transfer membrane (Amersham Life Sciences, Piscataway NJ). The membrane was then incubated with the specific antibody followed by the appropriate horseradish peroxidase-conjugated secondary antibody (Thermo Scientific, Pittsburgh, PA). The detection signal was developed with Amersham's enhanced chemiluminescence reagent and using FOTO/Analyst Luminary Work Station (Fotodyne Inc., Hartland, WI). The Western blots were quantified by densitometric analysis using Total lab Nonlinear Dynamic Image analysis software (Nonlinear USA, Inc., Durham, NC).