All lanes : HRP Anti-ATAD2 antibody [EPR12730] (ab201743)

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : ATAD2 knockout HAP1 whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 159 kDa


Exposure time: 20 minutes

ab201743 was shown to recognize ATAD2 in wild-type HAP1 cells as signal was lost at the expected MW in ATAD2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and ATAD2 knockout samples were subjected to SDS-PAGE. ab201743 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor^® 680) loading control) were incubated overnight at 4°C at 5000 dilution and 1/1000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

HRP Anti-ATAD2 antibody [EPR12730] (ab201743) at 1/5000 dilution + Saos 2 (Human epithelial-like osteosarcoma cell line) Whole Cell Lysate at 10 µg

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 159 kDa
Observed band size: 190 kDa why is the actual band size different from the predicted?


Exposure time: 1 minute

This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab201743 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.