||Rapid Technique for Estimating DNA Binding, Used to Evaluate 1-Nitropyrene Adduct Formation.
Jackson, M. A. ;
King, L. C. ;
Ball, L. M. ;
||Health Effects Research Lab., Research Triangle Park, NC. ;Northrop Services, Inc., Research Triangle Park, NC.
Deoxyribonucleic acids ;
Labeled substances ;
||Most EPA libraries have a fiche copy filed under the call number shown. Check with individual libraries about paper copy.
A simple and rapid procedure for estimating binding of radiolabelled material to DNA and protein is described. Protein was extracted from lysed rabbit alveolar macrophages with chloroform: iso-amyl-alcohol:phenol extraction. Nucleic acids were precipitated from the lysate, and hydrolysed with protease and NaOH to remove residual protein and RNA respectively. Bound radioactivity was quantitated by precipitation of DNA onto glass fiber filters. Protein labelled with H-leucine and DNA and RNA adducts formed from 1-nitro(c)pyrene by xanthine oxidase were used to define this procedure. C was shown to be bound to endogenous protein and DNA isolated from rabbit alveolar macrophages that had been incubated with 1-nitro(c)pyrene.