Inventors:
Max Costa - Ramsey NJ
Anatoly V. Zhitkovich - Greenwood NY
Assignee:
New York University - New York NY
International Classification:
C12Q 148
C12Q 168
C12Q 1533
Abstract:
Since sodium dodecyl sulfate (SDS) binds tightly to proteins but not to DNA, the addition of potassium chloride (KCl) to SDS lysates of cells, nuclei or mitochondria results in the formation of an insoluble precipitate which includes all proteins and detergent-resistant DNA-protein complexes separated from free DNA in the supernatant. The amount of SDS-precipitable DNA represents a measure of crosslinked DNA-protein. This precipitation method is useful for determining or quantitating crosslinked DNA-protein formed in cells, nuclei or mitochondria of animals following their exposure to crosslinking agents such as chromate, nickel, cis-Pt (II) diammine dichloride and formaldehyde. The method is also useful for evaluating agents for their ability to induce covalent DNA-protein crosslinking.