FIGURE 1.

Diagram showing preparatory steps for western blotting samples (S1 and P3), according to Danbolt et al. (1990). The tissue was homogenized in about 10 volumes of homogenization buffer and centrifuged (10 min, 1000 g, 4°C). The pellets (P1) were discarded. The combined supernatants (S1) were centrifuged again (20 min, 27000 g, 4°C), and the pellets (P2) were suspended in about 10 volumes of hypotonic buffer and centrifuged (20 min, 27000 g, 4°C). The pellets (P3) were resuspended in 10 volumes of homogenization buffer in order to eliminate the hypotonic buffer and centrifuged (20 min, 27000 g, 4°C). The final pellets (P3) were resuspended in appropiate volume.