Methods of use for and kits containing chemiluminescent compounds

A method for determining an analyte, said method comprising:

• a) combining (1) a medium suspected of containing said analyte and (2) a label reagent comprising a first specific binding pair (sbp) member associated with a chemiluminescent compound wherein said first sbp member is capable of binding to said analyte or to a second sbp member capable of binding to said analyte, to form a complex in an amount related to the presence of said analyte, said chemiluminescent compound having the formula:
• ##STR32## wherein:
• X" and Y" are linking groups each comprising a chain of one atom in length, said atom being independently selected from the group consisting of O, S and N wherein N is part of the functionality N (E).sub.p R", where E is selected from the group consisting of CO and SO.sub.2, p is an integer from 0-1, and R" is selected from the group consisting of H, lower alkyls and halogenated lower alkyls;
• Z" is a group linking X" and Y" comprising a chain of 1-2 carbon atoms in length;
• and where one or more hydrogens of said Y" and said Z" may be substituted by organic radicals;
• b) chemically activating said chemiluminescent compound;
• and
• c) detecting the amount of luminescence generated by said chemiluminescent compound, the amount thereof being related to the amount of analyte in said medium.

A method for determining an analyte, said method comprising:

• a) combining in an assay medium (1) a sample suspected of containing an analyte and (2) a label reagent comprising a member of a specific binding pair (sbp member) bound to a chemiluminescent compound under conditions wherein an sbp member complex involving said label reagent is formed in relation to the presence of analyte in said sample, said chemiluminescent compound having the formula:
• ##STR34## wherein:
• X"' and Y"' are linking groups each comprising one atom in length, said atom being independently selected from the group consisting of O, S and N wherein N is part of the functionality NH;
• Z"' is a group linking X"' and Y"' comprising a chain of 1-2 carbon atoms in length;
• and where one or more hydrogens of said Y"' and said Z"' may be substituted by organic radicals;
• b) chemically activating said chemiluminescent compound;
• and
• c) examining said assay medium for a signal, the presence or intensity thereof being related to the amount of analyte in said sample.

A method for determining an analyte, said method comprising:

• a) combining in an assay medium either wholly or partially sequentially (1) a sample suspected of containing an analyte, (2) a label reagent comprising a first member of a specific binding pair (sbp member) bound to a chemiluminescent compound and (3) an insolubilized reagent comprising a second sbp member under conditions wherein an sbp member complex involving said label reagent and said insolubilized reagent is formed in relation to the presence of analyte in said medium, said chemiluminescent compound having the formula:
• ##STR36## wherein:
• X' and Y' are linking groups each comprising a chain of one atom in length, said atom being independently selected from the group consisting of O, S, Se, and N wherein N is part of the functionality selected from the group consisting of NH, NR', NSO.sub.2 R' and NCOR', where R' is selected from the group consisting of alkyl, aryl and halogenated alkyl groups;
• Z' is a group linking X' and Y' comprising a chain of 1-2 carbon atoms in length;
• where one or more hydrogens of said Y' and said Z' may be replaced by one or more organic radicals which may be taken together to form rings or double bonds;
• b) separating said assay medium and said insolubilized reagent;
• c) chemically activating said chemiluminescent compound in said medium or on said insolubilized reagent;
• and
• d) examining said assay medium or said reagent for a signal, the presence or intensity thereof being related to the amount of analyte in said sample.