A rapid and sensitive method for determination of covalent binding of benzo[a]pyrene to proteins |
| |
Authors: | H Wallin C Schelin A Tunek B Jergil |
| |
Affiliation: | 1. Department of Chemical Carcinogenesis, Michigan Cancer Foundation, Detroit, MI U.S.A.;2. Division of Cancer Research, Department of Medicine, Michael Reese Hospital and Medical Center, Chicago, IL U.S.A. |
| |
Abstract: | A method is presented for the quantitative determination of covalent binding of metabolically activated benzo[a]pyrene to microsomal proteins. After incubation of radiolabelled benzo[a]pyrene with microsomes and NADPH, the mixture is applied to filter paper discs. These are immersed in ethanol to precipitate the proteins. Unbound radiolabel is removed by repeated washes of the filters in organic solvents before scintillation counting. The method is simple, rapid, sensitive and accurate, and works both with 14C- and 3H-labelled compounds. The method is suitable for measuring the incorporation of other radiolabelled xenobiotics to proteins of both microsomes and other subcellular fractions and for the analysis of binding to isolated proteins. |
| |
Keywords: | AAF 2-acetylaminofluorene ABP 4-aminobiphenyl ABP-GMP 3-phospho-4-aminobiphenyl AF 2-aminofluorene HPLC high performance liquid chromatography PDE bovine spleen phosphodiesterase II RNAase ribonuclease A TLC thin-layer chromatography TMS trimethylsilane |
本文献已被 ScienceDirect 等数据库收录! |