Recombinant HRP Anti-MIF antibody [EPR12463] (ab196645)

ab196645 was shown to recognize MIF (Macrophage migration inhibitory factor) in wild-type HAP1 cells as signal was lost at the expected MW in MIF knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and MIF knockout samples were subjected to SDS-PAGE. Ab196645 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 2 μg/ml and 1/20000 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 MES buffer system. The gel was run at 200V for 35 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 ab196645 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.