Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human pineal tissue labelling HECT E3 ubiquitin ligase with ab83853 at 1/100. A Cy3-conjugated donkey anti-rabbit IgG (1/200) was used as the secondary antibody. Positive staining shown in the cytoplasm of cell bodies and processes of pinealocytes. Magnification: 20X. Exposure time: 0.5 - 2.0 seconds. Left - DAPI. Middle - HECT E3 ubiquitin ligase. Right - Merge.
Anti-HECT E3 ubiquitin ligase antibody (ab83853) at 1 µg/ml (5% skim milk / PBS ) + HepG2 cell lysate at 10 µgSecondaryHRP conjugated anti-Rabbit IgG at 1/50000 dilution
ICC/IF image of ab83853 stained HeLa cells. The cells were 4% formaldehyde (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 (ab83853, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.