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Radial diffusion in gel for micro determination of enzymes

1972; Elsevier BV; Volume: 48; Issue: 1 Linguagem: Inglês

10.1016/0003-2697(72)90165-0

1096-0309

G. Schumacher, Wolf‐Bernhard Schill,

Advanced Proteomics Techniques and Applications

The principle of radial diffusion in substrate containing agar gel has been applied for quantitation of proteolytic activity in small volumes (2 μl). Urokinase, elastase, α-chymotrypsin, trypsin, and other proteases have been investigated. Clearing zones in the agar gel indicate the presence of the enzyme. A linear relationship was found between the logarithm of the proteinase concentrations and the diameters of the clearing zones for urokinase, elastin, and α-chymotrypsin. The trypsin standard curve represents a curved line in the semilogarithmic system. The reproducibility of these micro methods is satisfactory. They are suitable for the analysis of biological fluids, especially viscous secretions, and for screening of large numbers of chromatography fractions. The micro radial diffusion of enzymes in gel is a simple and time-saving method that does not require expensive equipment.