Immunohistochemistry analysis of paraffin-embedded human colon carcinoma tissue stained with BCAR3 antibody (ab65111) at 1/50 - 1/100 dilution, with (right) and without (left) the immunising peptide.
All lanes : Anti-BCAR3 antibody (ab65111) at 1/500 dilutionLane 1 : extracts from HeLa cellsLane 2 : extracts from Jurkat cellsLane 3 : extracts from HUVEC cellsLane 4 : extracts from HUVEC cells with immunising peptide at 5 µgLysates/proteins at 5 µg per lane.
ICC/IF image of ab65111 stained MCF7 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 (ab65111, 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.