Anti-MAD2 antibody (ab70383) at 1 µg/ml + NETN whole cell lysate from HeLa cells at 50 µgdeveloped using the ECL technique

ab70383 Immunoprecipitating MAD2 in human HeLa whole cell lysate. 200µg of cell lysate was incubated with primary antibody (2 µg/ml in NETN buffer) and matrix (Protein A/G) for 16 hours at 4°C. For western blotting ab70383 (1/1000) was used to confirm successful immunoprecipation.See Abreview

ICC/IF image of ab70383 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 (ab70383, 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.