All lanes : Anti-CDCA3 antibody (ab69608) at 1/500 dilutionLane 1 : extracts from HepG2 cellsLane 2 : extracts from HepG2 cells with immunising peptide at 5 µgLysates/proteins at 5 µg per lane.
Human colon carcinoma tissue stained with ab69608 at 1/50-1/100 dilution, with and without the immunising peptide.
ICC/IF image of ab69608 stained MCF7 cells. The cells were 4% PFA fixed (10mins) 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 (ab69608, 5µg/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.