Junctional Adhesion Molecule 1/JAM-A  contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted. The predicted molecular weight of Junctional Adhesion Molecule 1/JAM-A  is 32 kDa (SwissProt), however we expect to observe a banding pattern around 42 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 5% Bovine Serum Albumin before being incubated with ab125886 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.