A method for direct cross-linking of DNA bases containing aromatic amino groups to proteins.
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.
Conlon
K A
KA
Department of Pharmacological Sciences, University Medical Center at Stony Brook, State University of New York at Stony Brook, 11794-8651, USA.
Berrios
M
M
D000107:Acetylation; D000818:Animals; D002417:Cattle; D003432:Cross-Linking Reagents / Q000737:chemistry*; D004247:DNA / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D005821:Genetic Techniques; D006147:Guanine / Q000031:analogs & derivatives* / Q000737:chemistry; D015151:Immunoblotting; D012710:Serum Albumin, Bovine / Q000737:chemistry; D013387:Succinic Anhydrides / Q000737:chemistry
D003432:Cross-Linking Reagents; D013387:Succinic Anhydrides; D012710:Serum Albumin, Bovine; C024829:8-hydroxyguanine; D006147:Guanine; C031801:succinic anhydride; D004247:DNA