Anti-Bad antibody [Y208] (ab32445)

  Western blot - Bad antibody [Y208] (ab32445)

Anti-Bad antibody [Y208] (ab32445) at 1/5000 dilution + Hela cell lysate

Predicted band size : 18 kDa
Observed band size : 23 kDa (why is the actual band size different from the predicted?)

  Immunohistochemistry (Paraffin-embedded sections) - Bad antibody [Y208] (ab32445)

Immunohistochemical staining of paraffin-embedded human lymphoma using ab32445 at 1/100 dilution.

  Immunocytochemistry/ Immunofluorescence-Bad antibody [Y208](ab32445)

ICC/IF image of ab32445 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 (ab32445, 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.

  Flow Cytometry-Bad antibody [Y208](ab32445)

Overlay histogram showing MCF-7 cells stained with ab32445 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32445, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in MCF-7 cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.