초록 ▼

A support and method for the determination of enzyme activity in a biological extract is provided wherein the support comprises a fibrous material impregnated with a substrate and a water-soluble pH stabilizer comprising a first reagent suitable for a pH of enzyme activity over 7 or a second reagent

A support and method for the determination of enzyme activity in a biological extract is provided wherein the support comprises a fibrous material impregnated with a substrate and a water-soluble pH stabilizer comprising a first reagent suitable for a pH of enzyme activity over 7 or a second reagent suitable for a pH of enzyme activity of 7 or less; the viscose fibers are sufficiently loose in texture so as to be accessible both to products in solution and to tissuey and cellular mash or whole cells and sufficiently fine to permit adsorption and uniform distribution of reagents over the area thereof. The method comprises: (a) preparing a biological extract of the enzyme to be determined; (b) depositing a sample thereof on the support; (c) incubating said sample; and (d) placing on the support a developing reagent comprising a wetting agent and a buffer, and a coloring agent whereby the reactions are brought to a fixed pH, said pH being optimal for the colorimetric reaction between the substrate and the coloring agent.

대표청구항 ▼

A support for the determination of enzyme activity in a biological extract comprising a fibrous material impregnated with a substrate labeled with a naphthol and a buffer solution selected from the group consisting of a first buffer solution suitable for a pH of enzyme activity over 7 and a second b

A support for the determination of enzyme activity in a biological extract comprising a fibrous material impregnated with a substrate labeled with a naphthol and a buffer solution selected from the group consisting of a first buffer solution suitable for a pH of enzyme activity over 7 and a second buffer solution suitable for a pH of enzyme activity of 7 or less and free from color developing agents; said fibrous material being loose in texture so as to be accessible both to products in solution and to said biological extract consisting of tissuey and cellular mash or whole cells and sufficiently fine to permit adsorption and uniform distribution of reagents over the area thereof; and said naphthol substrate originally being colorless or pale yellow so that any subsequent coloration of the substrate, which results from coupling of said naphthol released therefrom with a diazonium salt color-developing agent to indicate the presence of enzyme activity, is readily apparent. An apparatus for determining enzyme activity in a biological extract comprising: (a) a tray having a plurality of cups contained therein; and (b) a plurality of discs located in the bottom of said cups, said discs being composed of a fibrous material impregnated with a substrate labeled with a naphthol and a buffer solution selected from the group consisting of a first buffer solution suitable for a pH of enzyme activity over 7 and a second buffer solution suitable for a pH of enzyme activity of 7 or less, and free from coloring agents; said fibrous material being loose in texture so as to be accessible both to products in solution and to biological samples of tissuey and cellular mash or whole cells and sufficiently fine to permit adsorption and uniform distribution of reagents over the area thereof and said naphthol substrate originally being colorless or pale yellow so that any subsequent coloration of the substrate, which results from coupling of the naphthol released therefrom with a diazonium salt color-developing agent to indicate the presence of enzyme activity, is readily apparent. A method of determining enzyme activity comprising: (a) preparing a biological extract of the enzyme whose activity is to be determined; (b) depositing a sample of said extract on a support comprising a fibrous material impregnated with a substrate labeled with a naphthol and a buffer solution selected from the group consisting of a first buffer solution suitable for a pH of enzyme activity over 7 and a second buffer solution suitable for a pH of enzyme activity of 7 or less, and free from color-developing agents; said fibrous material being loose in texture so as to be accessible both to products in solution and to biological samples of tissuey and cellular mash or whole cells and sufficiently fine to permit adsorption and uniform distribution of reagents over the area thereof; said substrate originally being colorless or pale yellow so that any subsequent coloration of the substrate, which results from coupling of the naphthol released therefrom with a color-developing agent to indicate the presence of enzyme activity, is readily apparent; (c) incubating said sample; (d) depositing on said support a developing reagent comprising a wetting agent, a buffer suitable for use at the desired pH and a diazonium salt color-developing agent for coupling with said released naphthol whereby the reactions are brought to a pH optimal for the colorimetric reaction between said substrate and said color-developing agent; (e) photochemically decomposing any unreacted color-developing agent; and (f) comparing said color reactions to a color scale indicative of the activity of the enzyme.