Figure 1

Characterization of the Eag1 antibodies. a. Western blot analysis of Eag1-expressing membrane preparations with anti-Eag1.62.mAb. A single protein is detected only when using large amounts of brain extract protein. b. Eag1.62.mAb stained CHO cells transfected with Eag1 (left) while cells transfected with Eag2 (right) show only faint background, indicating that Eag1.62.mAb does not recognise Eag2. c. CHO cells transiently expressing a chimera between EGFP and hEag1. Chimeras were stained with Eag1.62.mAb. The green fluorescence due to the presence of the chimera (upper left panel, marked EGFP) matches the staining pattern of the antibody (upper right panel, marked Eag1.62.mAb), as seen as yellow colour in the merged pseudo-colour image (lower panel, marked Overlay) Scale Bar: 20 μm. d-g. Light micrographs showing the immunohistochemical reaction for the monoclonal antibodies in rat hippocampus (d, Eag1.62.mAb; e, Eag1.33.mAb) and cerebellum (f, Eag1.62.mAb; g, Eag1.33.mAb), with (right column) and without (left column) pre-adsorption of the antibody to the corresponding fusion protein. The staining patterns are identical for both antibodies and are fully blocked by incubation with the corresponding epitopes, indicating that both antibodies specifically recognise the same molecular entity. Scale Bar: 50 μm.