HRP Anti-MIF antibody [EPR12463]

• WB

Lab

Western blot - HRP Anti-MIF antibody [EPR12463] (AB196645)

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.

All lanes:

Western blot - HRP Anti-MIF antibody [EPR12463] (ab196645) at 1/2000 dilution

Lane 1:

Y79 (Human retinoblastoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

Brain (Human) Tissue Lysate - fetal normal tissue (<a href='/en-us/products/unavailable/brain-human-tissue-lysate-fetal-normal-tissue-ab29467'>ab29467</a>) at 10 µg

Predicted band size: 12 kDa

Observed band size: 12 kDa

true

Exposure time: 4min

• WB

Lab

Western blot - 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.

All lanes:

Western blot - HRP Anti-MIF antibody [EPR12463] (ab196645) at 2 µg/mL

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

MIF knockout HAP1 whole cell lysate at 20 µg

Predicted band size: 12 kDa

Observed band size: 12 kDa

false