All lanes : Anti-Mammaglobin B antibody (ab53134) at 1/300 dilutionLane 1 : HepG2 cell extractLane 2 : HepG2 cell extract with immunising peptide
ab53134, at 1/50 dilution, staining Mammaglobin in paraffin embedded human breast carcinoma tissue by Immunohistochemistry in the absence (left image) or presence (right image) of the immunising peptide.
ICC/IF image of ab53134 stained HepG2 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 (ab53134, 5µg/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.