Detection of DNA Adducts in Human Breast Tissue
Abstract
The purpose of this project is to optimize and apply new methodology for the detection of unknown DNA adducts in breast and, for comparison, other human tissues. In this methodology the DNA adducts are labeled with a fluorescent dye to enhance their detection. Although the first dye tested (IMI1) was not fully stable, this problem was overcome by preparing IMI2. High dye Purity is required, and this was accomplished for IMI2 by means of flash chromatography/submarine gel electrophoresis, which brought the purity to 99.9998%. A basic protocol has been set up for the entire method, starting with human cell culture or human autopsy tissue samples, including breast samples and finishing with detection by capillary electrophoresis with laser induced fluorescence detection (CE-LIF). Standard adducts, after labeling with IMI2, were detected by matrix assisted laser desorption ionization mass spectrometry (MALDI-MS), with 300 fmol in the laser beam. Thus, the methodology is emerging as planned.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jul 01, 1998
- Accession Number
- ADB239319
Entities
People
- Roger W. Giese
Organizations
- Northeastern University