Anti-Presenilin 1/PS-1 antibody [5E12] - BSA and Azide free

• WB

Lab

Western blot - Anti-Presenilin 1/PS-1 antibody [5E12] - BSA and Azide free (AB255850)

This data was developed using 252856, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure time : 37 seconds

All lanes:

Western blot - Anti-Presenilin 1/PS-1 antibody [5E12] (<a href='/en-us/products/primary-antibodies/presenilin-1-ps-1-antibody-5e12-ab252856'>ab252856</a>) at 1/1000 dilution

All lanes:

HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/5000 dilution

Predicted band size: 53 kDa

Observed band size: 18 kDa

false

• WB

Lab

Western blot - Anti-Presenilin 1/PS-1 antibody [5E12] - BSA and Azide free (AB255850)

This data was developed using ab252856, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 3% NFDM/TBST.

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

Lanes 1-3 : Merged signal (red and green). Green - ab252856 observed at 18kDa. Red - loading control ab181602 (Rabbit monoclonal [EPR16891] to GAPDH) observed at 36 kDa.

Lanes 1-2 : ab252856 Anti-Presenilin 1/PS-1 antibody was shown to react with Presenilin 1/PS-1 in HAP1 cells in Western blot. Loss of signal was observed when Presenilin 1/PS-1 knockout sample was used. Wild-type and Presenilin 1/PS-1 knockout samples were subjected to SDS-PAGE.

ab252856 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated at 4℃ overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680CW) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800RD) preadsorbed (ab216772) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Presenilin 1/PS-1 antibody [5E12] (<a href='/en-us/products/primary-antibodies/presenilin-1-ps-1-antibody-5e12-ab252856'>ab252856</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 2:

PSEN1 knockout HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 3:

A431 (human epidermoid carcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rat IgG H&L (IRDye® 800CW ) (ab253031) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/20000 dilution

Predicted band size: 53 kDa

Observed band size: 18 kDa

false

• WB

Lab

Western blot - Anti-Presenilin 1/PS-1 antibody [5E12] - BSA and Azide free (AB255850)

This data was developed using the same antibody clone in a different buffer formulation (ab252856).

Western blot : Anti-PSEN1 antibody [5E12] (ab252856) staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab252856 was shown to bind specifically to PSEN1. A band was observed at 100 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-Rat IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Presenilin 1/PS-1 antibody [5E12] (<a href='/en-us/products/primary-antibodies/presenilin-1-ps-1-antibody-5e12-ab252856'>ab252856</a>) at 1/1000 dilution

Lane 1:

Wild-type PC-3 cell lysate at 20 µg

Lane 2:

PSEN1 knockout PC-3 cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 100 kDa

false