Myocilin, Abcam, ab41552

This antibody was included in a study aiming to characterize and validate commecially available antibodies raised against myocilin. Epitope mapping was conducted using recombinant myocilin subdomains cloned and expressed in E. coli. As positive control was used endogenously expressed full-length human myocilin secreted by trabecular meshwork (TM) cells.

We evaluated four antibodies (Ab41552, MABN866, MAB3446, sc-137233) for application in Western blotting (denaturing conditions) and ELISA (native protein).

Our data show that Ab41552 recognizes the far N-terminal residues 33-46 of myocilin. The antibody works well for WB, and detects E. coli expressed myocilin fragments containing these residues as well as the full-length human protein. However, prolonged exposure results in background bands in both sample types.

The epitope was confirmed in ELISA, which also showed a weak interaction with a myocilin fragment not containing the epitope, but with known high sequence similarity.

For further information see Patterson-Orazem et al., 2018 “Epitope mapping of commercial antibodies that detect myocilin” (www.ncbi.nlm.nih.gov/pubmed/29752947)