Anti-Bax antibody [6A7] (ab5714)

Immunocytochemistry/ Immunofluorescence - Bax antibody [6A7] (ab5714)

ab5714 staining human foreskin fibroblast cells by ICC/IF (Immunocytochemistry/immunoflurescence). 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, dilution 1/1000, was used as secondary antibody.

This image is courtesy of an Abreview submitted by Dr Alwin Scharstuhl

Immunocytochemistry/ Immunofluorescence - Bax antibody [6A7] (ab5714)

ab5714 staining Bax in human PMN cells by Immunocytochemistry/ Immunofluorescence. The cells were fixed in paraformaldehyde, permeabilised in 0.1% Triton X-100 and then blocked using 2% BSA for 1 hour at 22ºC. Samples were then incubated with primary antibody at 1/200 for 16 hours at 4ºC. The secondary antibody used was a goat anti-mouse IgG (H+L) conjugated to Alexa Fluor® 488 (green) used at a 1/500 dilution. Counterstained with DAPI (blue).

Image courtesy of an anonymous Abreview.

Flow Cytometry-Bax antibody [6A7](ab5714)

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 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. 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.

Western blot - Anti-Bax antibody [6A7] (ab5714)

Anti-Bax antibody [6A7] (ab5714) at 1/2000 dilution + whole tissue lysate prepared from human islet tissue at 20 µg

Secondary
Rabbit polyclonal Secondary Antibody to Mouse IgG - H&L (HRP) (ab6728) at 1/10000 dilution
developed using the ECL technique

Observed band size : 20 kDa (why is the actual band size different from the predicted?)
Additional bands at : 35 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 30 seconds

Image courtesy of an anonymous Abreview.