Recombinant HRP Anti-GSK3 beta antibody [Y174] (ab196911)

ab196911 was shown to recognize GSK3 beta in wild-type HAP1 cells as signal was lost at the expected MW in GSK3B knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and GSK3B knockout samples were subjected to SDS-PAGE. Ab196911 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor^® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/1000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

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 2% Bovine Serum Albumin before being incubated with ab196911 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.