ICC/IF image of ab20384 stained Mcf7 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 (ab20384, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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.

Ab20384 staining primary rat hepatocytes cells by EM. The cells were fixed and prepared by Tokuyasu method. The cryosections of 70nm were placed on 100 mesh grids. All the following steps were done at room temperature. Blocking was carried out for 30 min with 1% fish skin gelatine (FSG). The tissue sections were placed on a drop of 5 ul of first Ab (anti-LRP 8G1) for 20 min at dilutions of 1:400 or 1:600 seemed to work the best. After incubation with the primary antibody, 3. 5 x 2 min wash was done with PBS and the sections were placed on a drop of 5 ul of secondary Ab (goat anti-mouse) for 20 min (dilution 1:150 5. 5 x 2 min wash with PBS). Samples were placed on a drop of 5 ul of PAG10 for 15 min (dilution 1:60 or 1:70) and then 5 x 2 min wash with PBS. 2 min on 1% glutaraldehyde for fixation and 4 x 2.5 min on H2O. afterthat 1 second + 1 second + 6 min on drops of Methyl Cellulose/Uranyl Acetate (9:1) on ice. The surplus of MC/UA was removed , the slides were dried and obser