Anti-HSD17B4 antibody [2D3BB5BF10] (ab128565)

Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: HSD17B4 knockout  HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: A431 whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab128565 observed at 85 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab128565 was shown to specifically react with HSD17B4 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when HSD17B4 knockout cells were examined. Wild-type and HSD17B4 knockout samples were subjected to SDS-PAGE.  Ab128565 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

Immunofluorescent staining of HSD17B4 in HDFn cells (human) using ab128565.
The cells were paraformaldehyde fixed (4%, 20 min) and Triton X-100 permeabilized (0.1%, 15min). The cells were then incubated with the antibody (ab128565, 1µg/ml) for 2h at room temperature or over night at 4°C. The secondary antibody was (red) Alexa Fluor® 594 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. 10% Goat serum was used as the blocking agent for all blocking steps. The target protein locates to the peroxisome.