Fig. 1.
mAb 12D1 reacts with the LAH of HA2. (A) Hemagglutinin monomer. Amino acids 76–130 of HA2 are highlighted in red. (B) Lysates from 293T cells were transfected with pCAGGs-GFP or pCAGGs-GFP HA2 76–130, incubated with mAb 12D1, and pulled down with protein G beads. Pulled-down fractions were blotted with mAb 12D1 or rabbit anti-GFP. Arrows indicate location of the amino acid 76–130 region–GFP fusion protein. Anti-mouse HRP used to detect 12D1-binding reacts with the mouse Ig heavy and light chains within the pulled-down fraction. (C) Structural integrity of the 12D1 epitope within the LAH-KLH conjugate was confirmed by direct-binding ELISA. Hemagglutinin-specific mAb 36A7 does not bind within the amino acid 76–130 region of HA2.