The binding of anti-DNA antibodies as measured fluorometrically by ethidium bromide
Shepherd, J.D.; Fritzler, M.J.; Watson, J.I.; van de Sande, J.H.
Journal of Rheumatology 5(4): 391-398
1978
ISSN/ISBN: 0315-162X PMID: 84077 Document Number: 126401
The phenanthridine dye ethidium bromide (EB) intercalates with double-stranded DNA (dsDNA) resulting in an enhancement of fluorescence. Single-stranded DNA (ssDNA) does not show this fluorescent enhancement. Purified IgG from patients with Systemic Lupus Erythematosus (SLE) containing anti-dsDNA antibodies competes with EB for binding to DNA resulting in a decrease in fluorescence. This study has shown that antibodies which bind ssDNA in the Millipore filter radioimmunoassay displace EB from dsDNA showing that antigenic determinants are available for binding in the double-stranded molecule. This study introduces the EB assay and presents a comparison with the Millipore filter assay.