The Application of Polyacrylamide Gel Electrophoresis to the Characterization of Cholinesterase Enzymes Used in Detector Kits.
Abstract
Vertical disc gel electrophoresis on polyacrylamide gel tubes has been applied to the separation and identification of cholinesterases from commercially available sources. Cholinesterase from horse serum, bovine erythrocyte, and electric eel was fractionated using the Davis method for human serum proteins. The fractions were visualized with a protein stain and two cholinesterase substrates, indoxyl acetate and indoxyl butyrate. Each enzyme had a distinctive pattern of protein bands. The eel and bovine enzymes, which have high specific activities, showed fewer bands than did the horse enzyme which has lower specific activity (lower purity). All three enzymes hydrolyzed indoxyl acetate giving colored bands. Only the horse enzyme, however, gave colored bands with indoxyl butyrate, which is in agreement with the known substrate specificities of cholinesterases. The small sample size required, the wide variations possible in gel pore size, and buffer system composition permit optimization of sensitivity and resolution for the particular enzyme being investigated and make the method useful for analytical quality control, preparative scale separations, and characterization of small amounts of enzyme from intelligence sources.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 01, 1977
- Accession Number
- ADA035395
Entities
People
- Alan Goodman
- Hermann Martens
- Lawrence M. Mccormack