Anti-PERP antibody [EPR7885] - BSA and Azide free

• WB

Lab

Western blot - Anti-PERP antibody [EPR7885] - BSA and Azide free (AB248288)

False colour image of Western blot : Anti-PERP antibody [EPR7885] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab129083 was shown to bind specifically to PERP. A band was observed at 17 kDa in wild-type A431 cell lysates with no signal observed at this size in PERP knockout cell line ab270475 (knockout cell lysate ab270498). To generate this image, wild-type and PERP knockout A431 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 (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-PERP antibody [EPR7885] (<a href='/en-us/products/primary-antibodies/perp-antibody-epr7885-ab129083'>ab129083</a>) at 1/1000 dilution

Lane 1:

Wild-type A431 cell lysate at 20 µg

Lane 2:

PERP knockout A431 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

MOLT-4 cell lysate at 20 µg

Predicted band size: 21 kDa

Observed band size: 17 kDa

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• WB

Lab

Western blot - Anti-PERP antibody [EPR7885] - BSA and Azide free (AB248288)

This data was developed using the same antibody clone in a different buffer formulation (ab129083).

Lanes 1-3 : Merged signal (red and green). Green - ab129083 observed at 24 kDa. Red - loading control ab8245 observed at 36 kDa.

ab129083 Anti-PERP antibody [EPR7885] was shown to specifically react with PERP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265335 (knockout cell lysate ab258105) was used. Wild-type and PERP knockout samples were subjected to SDS-PAGE. ab129083 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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-PERP antibody [EPR7885] (<a href='/en-us/products/primary-antibodies/perp-antibody-epr7885-ab129083'>ab129083</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PERP knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PERP knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-perp-knockout-hela-cell-line-ab265335'>ab265335</a>)

Lane 3:

HaCaT 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: 21 kDa

Observed band size: 24 kDa

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