Recombinant Anti-Integrin alpha 4/CD49D antibody [EPR1355Y] (ab81280)

Lane 1: Integrin alpha 4 knockout HAP1 cell lysate (20 µg)

Lane 2: Wild-type HAP1 cell lysate (20 µg)

Lanes 1 and 2: Merged signal (red and green). Green - ab81280 observed at 150 kDa. Red - loading control, ab8226, observed at 42 kDa.

ab81280 was shown to recognize Integrin alpha 4 in wild-type HAP1 cells along with additional cross-reactive bands. No bands were observed when Integrin alpha 4 knockout samples were used. Wild-type and Integrin alpha 4 knockout samples were subjected to SDS-PAGE. ab81280 and ab8226 (loading control to beta actin) were diluted 1/2000 and 1/1000 respectively and incubated overnight at 4°C. 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 1/10,000 dilution for 1hr at room temperature before imaging.

Lanes 1 - 2: Merged signal (red and green). Green - ab81280 observed at 150 kDa. Red - loading control, ab8245, observed at 37 kDa.

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab81280 and ab8245 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at a 1:10000 dilution for 1hr at room temperature and then imaged.