Anti-Presenilin 1/PS-1 抗体 [EP2000Y] (ab76083)

Western blot: Anti-PSEN1 antibody [EP2000Y] (ab76083) staining at 1/2500 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab76083 was shown to bind specifically to PSEN1. A band was observed at 18 kDa in wild-type PC-3 cell lysates with no signal observed at this size in PSEN1 knockout cell line. To generate this image, wild-type and PSEN1 knockout PC-3 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Presenilin 1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HEK293 whole cell lysate (20 µg)
Lane 4: HeLa whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab76083 observed at 18 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab76083 was shown to recognize Presenilin 1 in wild-type HAP1 cells as signal was lost at the expected MW in Presenilin 1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Presenilin 1 knockout samples were subjected to SDS-PAGE. Ab76083 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 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 1/10000 dilution for 1 hour at room temperature before imaging.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human glioma tissue sections labeling Presenilin 1/PS-1 with Purified ab76083 at 1:100 dilution (1.31 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

The 18 KD band is the C-terminal fragment after proteolytic process reported by PMID 9173929. But we are not sure how to define the extra band around 75 KD. 

The 18 KD band is the C-terminal fragment after proteolytic process reported by PMID 9173929.