All lanes : Anti-PKC zeta antibody (ab51157) at 1/500 dilutionLane 1 : extracts from 3T3 cells treated with 125ng/ml PMA for 30min, without immunizing peptideLane 2 : extracts from 3T3 cells treated with 125ng/ml PMA for 30min, with immunizing peptide
Immunohistochemical analysis of PKC zeta expression in paraffin-embedded human lung carcinoma tissue using 1/50 ab51157. Left: untreated sample. Right: sample treated with immunizing peptide (negative control).
ICC/IF image of ab51157 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab51157, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Anti-PKC zeta antibody (ab51157) at 1/1000 dilution + Active human PKC zeta full length protein (ab60848) at 0.01 µgSecondaryGoat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.Exposure time : 1 minute