(A) Ln-332 is composed of α3, β3, and γ2 chains assembled into α3β3γ2 heterotrimer via a three-s tranded α-helical coiled-coil domain. A globular domain (G domain) uniquely exists at the C-terminal region of the α3 chain, which consists of five tandem laminin G-like (LG) repeats module (LG1-LG5). The linker region between LG3 and LG4 (indicated by arrowhead) is the target for processing by serine proteases such as plasmin. (B) SDS-PAGE of Ln-332. Purified Ln-332 was run on 4–12 % gradient gel under reducing conditions. Separated Ln-332 chains were stained with Coomassie Brilliant Blue. The gel includes bands identified as the α3 (~150 kDa), β3 (~135 kDa), and γ2 (~140 kDa and ~80 kDa) chains. (C) Immunoblot analysis was carried out against Ln-332 chains using CM6, anti-β3, anti-γ2 antibody (TR1). CM6, anti-β3 pAb, and TR1 were used as a first antibody at the diluted ratio of 1: 100, 1: 1000, 1: 200, respectively. Anti-rabbit or mouse HRP conjugated IgG was used as a secondary antibody at the diluted ratio of 1: 1000. These results suggest that CM6 recognizes the α3 chain, but not β3 and γ2 chains, of Ln-332.