A blotting method for monitoring the formation of chemically induced DNA-protein complexes.
The formation and identification of DNA-protein crosslinks are usually detected by filter binding assays such as alkaline elution. We describe a modified blotting method to selectively identify DNA-protein complexes (DPCs) formed in vitro by either Cr3+ ion or formaldehyde. This protocol allows DPC formation in vitro to be assayed with various chemical agents, requires minimal usage of radioactivity, and is performed in a shorter time frame than that commonly used to resolve DPCs from free proteins and unbound DNA.
Cohen
M D
MD
Department of Environmental Medicine, New York University Medical Center, New York 10016.
Miller
C A
CA
Xu
L S
LS
Snow
E T
ET
Costa
M
M
D000199:Actins; D000418:Albumins; D000818:Animals; D015153:Blotting, Western / Q000379:methods*; D002417:Cattle; D002857:Chromium; D003432:Cross-Linking Reagents; D004247:DNA; D004268:DNA-Binding Proteins; D005557:Formaldehyde; D006657:Histones; D066298:In Vitro Techniques
D000199:Actins; D000418:Albumins; D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; D006657:Histones; D002857:Chromium; D005557:Formaldehyde; D004247:DNA