Fig. 4

The graph represents the recognition of multispecies sera by the SA-TRIM21-HRP protein. Animal sera (50 ng) were coated onto polystyrene plates, blocked with 1%BSA, incubated with SA-TRIM21 (0.5 μg/well), and biotinylated HRP (1:2500). Control experiments were performed using immobilized BSA into wells. After washing with PBST (low salt), 50 μL chromogenic substrate TMB was added to the wells, incubated for 10 min, followed by the addition of 50 μL H2SO4 to quench the reaction followed by OD₄₅₀ nm reading using an ELISA plate reader. The data shows the ratio of serum/control OD ₄₅₀ reading. Experiments were performed in triplicates, and error bars represent standard deviation