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Anti-MAD2 antibody (ab70383) at 1 µg/ml + NETN whole cell lysate from HeLa cells at 50 µg
developed using the ECL technique
Predicted band size : 24 kDa
Observed band size : 24 kDa
Additional bands at : 38 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 1 minute
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 4ºC. For western blotting ab70383 (1/1000) was used to confirm successful immunoprecipation.
This image is courtesy of an anonymous 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 +4ºC. 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.
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