Anti-Presenilin 1/PS-1 抗体 [EP2000Y]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Presenilin 1/PS-1 antibody [EP2000Y] (AB76083)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Presenilin 1/PS-1 antibody [EP2000Y] (AB76083)

Immunohistochemical analysis of paraffin-embedded human cerebral cortex and human Alzheimer's brain tissue* labelling PSEN1 with ab76083 at 1ug/ml followed by a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Positive staining in human cerebral cortex and negative staining in Alzheimer's brain.

The section was incubated with ab76083 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0 epitope retrieval solution1) for 20 mins.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• WB

Lab

Western blot - Anti-Presenilin 1/PS-1 antibody [EP2000Y] (AB76083)

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.

Lane 1:

Western blot - Anti-Presenilin 1/PS-1 antibody [EP2000Y] (ab76083)

Lanes 2 - 4:

Western blot - Anti-Presenilin 1/PS-1 antibody [EP2000Y] - BSA and Azide free (<a href='/products/primary-antibodies/presenilin-1-ps-1-antibody-ep2000y-bsa-and-azide-free-ab227985'>ab227985</a>) at 1/5000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

Presenilin 1/PS-1 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HEK293 whole cell lysate at 20 µg

Lane 4:

HeLa whole cell lysate at 20 µg

Predicted band size: 53 kDa

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• WB

Lab

Western blot - Anti-Presenilin 1/PS-1 antibody [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.

All lanes:

Western blot - Anti-Presenilin 1/PS-1 antibody [EP2000Y] (ab76083) at 1/2500 dilution

Lane 1:

Wild-type PC-3 cell lysate at 20 µg

Lane 2:

PSEN1 knockout PC-3 cell lysate at 20 µg

Lane 3:

Wild-type HAP1 cell lysate at 20 µg

Lane 4:

PSEN1 knockout HAP1 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

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• WB

Lab

Western blot - Anti-Presenilin 1/PS-1 antibody [EP2000Y] (AB76083)

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

All lanes:

Western blot - Anti-Presenilin 1/PS-1 antibody [EP2000Y] (ab76083) at 1/1000 dilution

All lanes:

Human fetal brain lysates at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 53 kDa

Observed band size: 18 kDa

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• WB

Lab

Western blot - Anti-Presenilin 1/PS-1 antibody [EP2000Y] (AB76083)

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.

All lanes:

Western blot - Anti-Presenilin 1/PS-1 antibody [EP2000Y] (ab76083) at 1/1000 dilution

Lane 1:

Mouse brain lysates at 15 µg

Lane 2:

Rat brain lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 53 kDa

Observed band size: 18 kDa

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