Fig. 3.

Determination of mAb150 reactive epitope. A-i. Coomassie blue stained SDS-PAGE protein gel of recombinant protein lysates of Ax1, Ax2 and Ax3, A-ii. Immunoblotting of Ax1, Ax2 and Ax3 recombinant lysates probed with mAb150 and consequently with Anti-GST (A-iii); B-i. Immunoblotting of Ax4 and Ax5 recombinant lysates probed with mAb150 and consequently with Anti-GST (B-ii); C. Coomassie blue stained SDS-PAGE protein gel of recombinant protein lysates of Ax4, Ax5 and Ax6 (left panel) and immunoprecipitated with mAb150 (right panel), followed by immunoblotting with mAb150 (D-i) and Anti-Anxa2 (D-ii); E. Immunoblotting of recombinant protein lysates of Ax7 and Ax8 with mAb150 (E-i) and Anti-GST(E-ii); F. ELISA-based profiling of mAb150 reactivity with Ax4, Ax5, Ax7 and Ax8 fragments; G. ELISA-based profiling of reactivity with synthetic peptides Ap1-Ap12 of (G-i) mAb150, and (G-ii) commercial AnxA2 C-terminal antibody (sc-166,762); H-i. Mapping of mAb150 epitope on AnxA2 1–50 aa sequence by 5 different servers, indicating (H-ii) overlapping epitope region (annotated by different color lines) in proximity of the N-terminal region (a core overlapping epitope is highlighted by gray background, annotated as *), (H-iii). Molecular surface view model that indicates its location (marked in pink) on the AnxA2 protein (green).