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A sensitive microplate assay for the detection of proteolytic enzymes using radiolabeled gelatin
B D Robertson1, G E Kwan-Lim, R M Maizels
1Department of Pure and Applied Biology, Imperial College of Science and Technology, London, United Kingdom.
Analytical Biochemistry
|July 1, 1988
Abstract:
A sensitive, microplate assay is described for the detection of a wide range of proteolytic enzymes, using radio-iodine-labeled gelatin as substrate. The technique uses the Bolton-Hunter reagent to label the substrate, which is then coated onto the wells of polyvinyl chloride microtiter plates. By measuring the radioactivity released the assay is able to detect elastase, trypsin, and collagenase in concentrations of 1 ng/ml or less, while the microtiter format permits multiple sample handling and minimizes sample volumes required for analysis.