Mouse anti domain 6 antibody

Wells were coated with increasing concentrations of tropoelastin at 4 °C overnight and washed with PBS to remove unbound tropoelastin. Wells were blocked with 3% (w/v) BSA for 1 h. Bound tropoelastin was separately detected with one of three primary antibodies as follows: (a) 1:2000 BA4 mouse anti-elastin antibody (Sigma); (b) 1:500 rabbit anti-C-terminal antibody (custom-made by Biomatik); or (c) 1:5000 mouse anti-domain 6 antibody (custom-made by AbMart). Wells were washed and incubated with 1:5000 goat anti-mouse or anti-rabbit IgG conjugated with horseradish peroxidase for 1 h. Wells were visualized with ABTS solution (1.04 mg/ml 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid, 0.05% (v/v) H₂O₂, 10 mm CH₃COONa, 5 mm Na₂HPO₄) at 37 °C for 1 h, and absorbance readings were measured at 405 nm.