Immunocytochemistry/ Immunofluorescence - Anti-Bax antibody [6A7] (ab5714)This image is courtesy of an Abreview submitted by Dr Alwin Scharstuhl
Immunofluorescence analysis of Human foreskin fibroblast cells, staining Bax with ab5714.
Cells were fixed with paraformaldehyde, permeabilized with 0.1% saponin prior to being blocked in 1% BSA + 2% normal goat serum for 30 mins at 20°C. Samples were incubated with 5 µg/ml primary antibody for 45 mins at 20°C; the diution buffer was 1% BSA, 0.1% saponin, 0.05% NaN3 in PBS. An Alexa Fluor® 594-conjugated Goat polyclonal to mouse IgG (ab150116), dilution 1/1000, was used as secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-Bax antibody [6A7] (ab5714)This image is courtesy of an anonymous Abreview
Immunofluorescence analysis of Human PMN cells staining Bax with ab5714.
The cells were fixed in paraformaldehyde, permeabilised in 0.1% Triton X-100 and then blocked using 2% BSA for 1 hour at 22°C. Samples were then incubated with primary antibody at 1/200 for 16 hours at 4°C. The secondary antibody used was a goat anti-mouse IgG (H+L) conjugated to Alexa Fluor® 488 (green) (ab150113) used at a 1/500 dilution. Counterstained with DAPI (blue).
Overlay histogram showing HeLa cells stained with ab5714 (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 (ab5714, 1/10 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.