8-hydroxyguanine and succinic-anhydride

A one step method to cross-link DNA bases containing aromatic amino groups directly to proteins was developed. No chemical modification of the base is required prior to conjugation, which is performed at neutral pH. Work focused on 8-oxoguanine and the carrier protein, bovine serum albumin. Conjugates were stable after sodium dodecyl sulfate (SDS)-induced protein denaturation and were characterized by UV spectroscopy, enzyme linked immunosorbent assay (ELISA), SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot analyses. This method is a viable alternative to existing procedures for generating DNA base-protein conjugates for antibody characterization and affinity purification.

Topics: Acetylation; Animals; Cattle; Cross-Linking Reagents; DNA; Electrophoresis, Polyacrylamide Gel; Genetic Techniques; Guanine; Immunoblotting; Serum Albumin, Bovine; Succinic Anhydrides