Western blot analysis of TTR antibody was performed by loading 1000 ng, 500 ng, 250 ng, 125 ng, 62.5 ng and 31.25 ng of a Human Transthyretin Recombinant protein, 1000 ng and 500 ng of BSA and 5 µl PageRuler Plus Prestained Protein Ladder (Product # 26619) onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Fast Blotter (Product # 62288) and blocked with 5% milk in TBST for at least 1 hour at room temperature. Transthyretin was detected at ~46 kD using a TTR mouse monoclonal antibody (Product # MA1-221) at a concentration of 1 µg/mL in 5% milk in TBST overnight at 4°C on a rocking platform, followed by an incubation with a goat anti-mouse IgG-HRP Superclonal secondary antibody (Product # A28177) at a dilution of 1:2000 for at least 1 hour at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078).