Figure 3. MAb 54-1 binds brachyury with a high affinity.

(A) Comparison between MAb 54-1 and commercial anti-brachyury antibodies (H-210 and ab57480) by ELISA. (B) Western blot analysis of endogenously expressed brachyury in three chordoma cell lines using the indicated antibodies. Bottom panels: Level of actin-normalized protein for each cell line as detected with the indicated antibodies, corresponding to the full length (49 KDa) and the shorter (45 KDa) protein band. (C) Western blot of protein lysates from H460 Con shRNA vs. Br shRNA utilizing the H-210 polyclonal Ab. (D) Western blot of PANC-1 cells expressing a control plasmid (pCMV), a shorter isoform (pBr-Short), or the full-length brachyury protein (pBr) using MAb 54-1.