Specific Radioisotopic Assay for Cholinesterase
Abstract
The radiometric method (I) for measuring ChE activity (Siakotos et al., Biochem. Med. 3, 1, 1969) was modified to preclude the use of p-dioxane, a hazardous material. The modified procedure (II) uses 0.4 N perchloric acid (PCA) , instead of p-dioxane, to denature the ChE and stop hydrolysis of 14C- acetylcholine (ACh). The unreacted substrate (ACh) is removed by cationic exchange resin suspended in water. The supernatant (acidic water solution) containing the product of hydrolysis, 14C-acetic acid, is mixed with nonhazardous scintillation cocktail and counted. The incubation mixture (37 degrees C) for II is similar to I and is composed of 0.1 ml of buffer-salt solution (pH 7.8), 0.1 ml of guinea pig whole blood (WB)-water suspension and 0. 1 ml of 3mM ACh solution. Keywords: Acetylcholinesterase; Acetylcholine; Radiometric ChE assay; Physostigmine; Irreversible ChE inhibitors; Titrametric ChE assay.
Document Details
- Document Type
- Technical Report
- Publication Date
- May 01, 1990
- Accession Number
- ADA224444
Entities
People
- Brian G. Talbot
- Dana R. Anderson
- Larrel W. Harris
- Willard J. Lennox
Organizations
- United States Army Medical Research Institute of Chemical Defense