Indirect ELISA and competitive ELISA results show that this antibody is specific to ac4C (N4-acetylcytidine). Indirect ELISA (blue curve, refer to top X-right Y axis) was performed by coating BSA conjugated ac4C at 50ng/well followed by blocking with 5% non fat milk. 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 Ac4C or its structure analogue compounds was mixed in 20ng/mL primary antibody (red curve, refer to bottom X-left Y axis). This data was developed using the same antibody clone with 68498-1-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 5% milk and blotted with N4-acetylcytidine antibody 68498-1-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 Monoclonal antibody 66360-1-Ig at the same dose. This data was developed using the same antibody clone with 68498-1-PBS in a different storage buffer formulation.