Premium
A FILTER BINDING ASSAY FOR CHARACTERIZATION OF ANTIBODIES TOWARDS DAMAGED DNA
Author(s) -
Helland Dag E.,
Eggset Guri,
Krokan Hans
Publication year - 1987
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/j.1751-1097.1987.tb05367.x
Subject(s) - polyclonal antibodies , dna , antibody , microbiology and biotechnology , nitrocellulose , chemistry , dissociation constant , binding constant , binding site , biology , biochemistry , immunology , receptor , membrane
A nitrocellulose filter binding assay was used to characterize rabbit antibodies specific for UV‐irradiated DNA. The dissociation constant for the UV‐DNA‐antibody complex was found to be 4.2 × 10 −12 M. No significant binding to unirradiated DNA was observed. Unlabelled single‐stranded and double‐stranded DNA competed with equal efficiency for the radioactively labelled antigen (UV‐irradiated φX174 DNA). The antibodies also bound to OsO 4 ‐treated DNA, suggesting that these polyclonal antibodies also recognize minor photoproducts. Caffeine efficiently decreased binding of the antibodies to UV‐irradiated DNA.