Indirect ELISA and competitive ELISA results show that this antibody is specific to Pseudouridine. Indirect ELISA was performed by coating BSA conjugated Pseudouridine at 10ng/well followed by blocking with 1% BSA. Serial diluted primary antibody was added to the plates and incubated at 37℃. HRP-goat anti-mouse was used for detection. Competitive ELISA was performed similarly except that different concentration of Pseudouridine or its structure analogue compounds are mixed in primary antibody (fixed at 10 ng/mL). This data was developed using the same antibody clone with 68578-1-PBS in a different storage buffer formulation.

Total RNA was isolated from HeLa cell line and was dotted to NC membrane at different amount as indicated above the dots. The membrane was blocked with BSA and blotted with Pseudouridine antibody 68578-1-Ig at 1:2000 followed by incubation of HRP-goat anti-mouse secondary antibody. Signal was developed by ECL substrate. A parallel dot blot was performed using mouse IgG1 isotype control antibody 66360-1-Ig at the same dose. This data was developed using the same antibody clone with 68578-1-PBS in a different storage buffer formulation.