Anti-Paxillin antibody [M107]

• WB

Unknown

Western blot - Anti-Paxillin antibody [M107] (AB23510)

Membrane was incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.

Lanes 1 - 2:

Western blot - Anti-Paxillin antibody [M107] (ab23510)

Lanes 3 - 4:

Anti-Paxillin (Ser178) antibody

Lanes 1 and 3:

A431 cells (20 ug/lane) serum starved overnight

Lanes 2 and 4:

A431 cells (20 ug/lane) serum treated with EGF (100 ng/ml) for 5 min.

Predicted band size: 65 kDa

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• WB

Lab

Western blot - Anti-Paxillin antibody [M107] (AB23510)

Lanes 1 - 4 : Merged signal (red and green). Green - ab23510 observed at 65 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab23510 was shown to recognize PXN in wild-type A-431 cells as signal was lost at the expected MW in PXN knockout cell line ab261892 (knockout cell lysate ab261701). Additional cross-reactive bands were observed in the wild-type and knockout samples. Wild-type and PXN knockout samples were subjected to SDS-PAGE. ab23510 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Paxillin antibody [M107] (ab23510) at 1/1000 dilution

Lane 1:

Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

PXN knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human PXN (Paxillin) knockout A-431 cell line (<a href='/en-us/products/cell-lines/human-pxn-paxillin-knockout-a-431-cell-line-ab261892'>ab261892</a>)

Lane 3:

U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate at 20 µg

Lane 4:

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

Predicted band size: 65 kDa

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