Abstract: A method for determining the presence of an analyte which is a specific binding pair member in a sample suspected of containing the analyte is described. The method involves: bringing together in an aqueous medium, the sample, an enzyme bound to a first specific binding pair member and an inhibitor for the enzyme bound to a second specific binding pair member wherein each specific binding pair member is capable of binding to the analyte or to a specific binding pair member complementary to the analyte; analyzing the medium for the activity of said enzyme; and relating the activity to the amount of analyte present in the medium. Compositions of matter and kits are also described.

Abstract: A method for determining the presence of an analyte which is a specific binding pair member in a sample suspected of containing the analyte is described. The method involves: bringing together in an aqueous medium, the sample, an enzyme bound to a first specific binding pair member and an inhibitor for the enzyme bound to a second specific binding pair member wherein each specific binding pair member is capable of binding to the analyte or to a specific binding pair member complementary to the analyte; analyzing the medium for the activity of said enzyme; and relating the activity to the amount of analyte present in the medium. Compositions of matter and kits are also described.

Abstract: A method is disclosed for stabilizing a conjugate of an enzyme and a member of a specific binding pair (enzyme conjugate). The method comprises the step of combining the enzyme conjugate with an effective amount of an antibody for the enzyme where the antibody does not substantially inhibit the activity of the enzyme. The invention has application to assays for the determination of an analyte wherein enzyme conjugates are employed. The improvement comprises employing as a reagent in the assay an immune complex of an enzyme conjugate and an antibody for the enzyme where the antibody does not substantially inhibit the activity of the enzyme. Compositions comprising such an immune complex and kits comprising such an immune complex in packaged combination with other assay reagents are also disclosed.

Abstract: Methods are disclosed for conducting assays. One such method comprises providing in combination a first bibulous member zone ("first zone") and a liquid medium containing a component. The first zone has non-diffusively bound thereto a reagent interreactive with the component. Conditions are selected wherein the liquid medium and at least a portion of the component contained therein traverse all of the first zone and migrate by capillary migration into a second bibulous member zone ("second zone"). The second zone is of a different composition than the first zone and is incapable of specifically binding the component except when an analyte is to be detected and the method further includes causing a reagent to become bound to the first bibulous member zone in relation to the amount of analyte present.

Abstract: A method is disclosed for conducting an assay for an analyte. The method comprises causing a specific binding pair member in a first aqueous medium to become bound to a first bibulous member by contacting a portion of the first bibulous member with the medium. The first bibulous member is in liquid receiving relationship with an absorbent member. The contacting is carried out under conditions wherein the first medium traverses the first bibulous member and at least a portion of the absorbent member by capillary action. The method further comprises causing the first bibulous member to come into liquid receiving relationship with a second bibulous member. A reagent in a second aqueous medium is absorbed by and preferably becomes non-diffusively bound to the second bibulous member in relation to the presence of analyte in the first medium.

Abstract: Methods are disclosed for conducting assays. One such method comprises providing in combination a first bibulous member zone ("first zone") and a liquid medium containing a component. The first zone has non-diffusively bound thereto a reagent interreactive with the component. Conditions are selected wherein the liquid medium and at least a portion of the component contained therein traverse all of the first zone and migrate by capillary migration into a second bibulous member zone ("second zone"). The second zone is of a different composition than the first zone and is incapable of specifically binding the component except when an analyte is to be detected and the method further includes causing a reagent to become bound to the first bibulous member zone in relation to the amount of analyte present.

Abstract: Methods are disclosed for conducting assays. One such method comprises providing in combination a first bibulous member zone ("first zone") and a liquid medium containing a component. The first zone has non-diffusively bound thereto a reagent interreactive with the component. Conditions are selected wherein the liquid medium and at least a portion of the component contained therein traverse all of the first zone and migrate by capillary migration into a second bibulous member zone ("second zone"). The second zone is of a different composition than the first zone and is incapable of specifically binding the component except when an analyte is to be detected and the method further includes causing a reagent to become bound to the first bibulous member zone in relation to the amount of analyte present.

Abstract: A method is disclosed for conducting an assay for an analyte. The method comprises causing a specific binding pair member in a first aqueous medium to become bound to a first bibulous member by contacting a portion of the first bibulous member with the medium. The first bibulous member is in liquid receiving relationship with an absorbent member. The contacting is carried out under conditions wherein the first medium traverses the first bibulous member and at least a portion of the absorbent member by capillary action. The method further comprises causing the first bibulous member to come into liquid receiving relationship with a second bibulous member. A reagent in a second aqueous medium is absorbed by and preferably becomes non-diffusively bound to the second bibulous member in relation to the presence of analyte in the first medium.

Abstract: Methods are disclosed for conducting assays. One such method comprises providing in combination a first bibulous member zone ("first zone") and a liquid medium containing a component. The first zone has non-diffusively bound thereto a reagent interreactive with the component. Conditions are selected wherein the liquid medium and at least a portion of the component contained therein traverse all of the first zone and migrate by capillary migration into a second bibulous member zone ("second zone"). The second zone is of a different composition than the first zone and is incapable of specifically binding the component except when an analyte is to be detected and the method further includes causing a reagent to become bound to the first bibulous member zone in relation to the amount of analyte present.

Abstract: A method is disclosed for conducting an assay for an analyte. The method comprises causing a specific binding pair member in a first aqueous medium to become bound to a first bibulous member by contacting a portion of the first bibulous member with the medium. The first bibulous member is in liquid receiving relationship with an absorbent member. The contacting is carried out under conditions wherein the first medium traverses the first bibulous member and at least a portion of the absorbent member by capillary action. The method further comprises causing the first bibulous member to come into liquid receiving relationship with a second bibulous member. A reagent in a second aqueous medium is absorbed by and preferably becomes a non-diffusively bound to the second bibulous member in relation to the presence of analyte in the first medium.

Abstract: Macromolecular compositions are provided which find use as glycosidase substrates, particularly where steric modulation of enzyme activity is involved. The compositions have a macromolecular hub to which is linked by means of a spacer arm a chromophore, which in turn is bonded to a sugar by an enzymatically labile glycosidyl linkage. The chromophore is chosen so as to undergo a substantial change in its chromophoric properties upon cleavage of the glycosidyl linkage. The enzyme activity can be determined spectrophotometrically due to the change in the absorptive or fluorescent properties of the chromophore upon cleavage of the glycosidyl linkage. Varying turnover rates for the enzyme are observed depending upon the nature of the macromolecular hub, the spacer arm, and the chromophore.