All lanes : Anti-SAE2 / UBA2 antibody (ab58451) at 1/500 dilutionLane 1 : extracts from 293 cells treated with UV (5mins)Lane 2 : extracts from 293 cells treated with UV (5mins) with immunizing peptide
Ab58451 at 1/50-1/100 dilution staining human lung carcinoma without and with blocking peptide; paraffin embedded.
ICC/IF image of ab58451 stained Hek293 cells. The cells were 100% methanol fixed (5 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 (ab58451, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Alexa Fluor® 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.