Anti-PICALM antibody [EPR12177] - BSA and Azide free

• WB

Lab

Western blot - Anti-PICALM antibody [EPR12177] - BSA and Azide free (AB249736)

This data was developed using the same antibody clone in a different buffer formulation (ab172962).

Lanes 1 - 4 : Merged signal (red and green). Green - ab172962 observed at 70-75 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

ab172962 was shown to react with PICALM in U-2 OS wild-type cells in Western blot. Loss of signal was observed when PICALM knockout sample was used. U-2 OS wild-type and PICALM knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween^®) before incubation with ab172962 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 5000 dilution and a 1 in 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 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PICALM antibody [EPR12177] (<a href='/en-us/products/primary-antibodies/picalm-antibody-epr12177-ab172962'>ab172962</a>) at 1/5000 dilution

Lane 1:

Wild-type U-2 OS whole cell lysate at 20 µg

Lane 2:

PICALM knockout U-2 OS whole cell lysate at 20 µg

Lane 2:

Western blot - Human PICALM knockout U-2 OS cell line (<a href='/en-us/products/cell-lines/human-picalm-knockout-u-2-os-cell-line-ab262500'>ab262500</a>)

Lane 3:

PC3 (Human prostate adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 4:

Caco-2 (Human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 70 kDa

Observed band size: 70-75 kDa

false

• WB

Lab

Western blot - Anti-PICALM antibody [EPR12177] - BSA and Azide free (AB249736)

This data was developed using the same antibody clone in a different buffer formulation (ab172962).

Lanes 1-3 : Merged signal (red and green). Green - ab172962 observed at 70-75 kDa. Red - loading control ab8245 observed at 36 kDa.

ab172962 Anti-PICALM antibody [EPR12177] was shown to specifically react with PICALM in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265653 (knockout cell lysate ab258113) was used. Wild-type and PICALM knockout samples were subjected to SDS-PAGE. ab172962 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PICALM antibody [EPR12177] (<a href='/en-us/products/primary-antibodies/picalm-antibody-epr12177-ab172962'>ab172962</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PICALM knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PICALM knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-picalm-knockout-hela-cell-line-ab265653'>ab265653</a>)

Lane 3:

Caco-2 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 70 kDa

Observed band size: 70-75 kDa

false