Protein proximity assay in formalin fixed paraffin embedded tissue using caged haptens

A compound having Formula (IE) or (IF):

• wherein
• “Hapten” is a pyrazole, a nitropyrazole, a benzofuran, an oxazole, an oxazole sulfonamide, a thiazole, or a thiazole sulfonamide;
• Y is selected from the group consisting of a carbonyl-reactive group, an amine-reactive group, and a thiol-reactive group, wherein the carbonyl-reactive group is selected from the group consisting of a hydrazine and an amine, wherein the amine-reactive group is a NHS ester, a sulfo-NHS, an isothiocyanate, an isocyanate, an acyl azide, a sulfonyl chloride, an aldehyde, a glyoxal, an epoxide, a oxirane, a carbonate, an aryl halide, an imidoester, or an anhydrides, and wherein the thiol-reactive group is a haloacetyl group, an alkyl halide, a maleimide, an aziridine, an acryloyl, a vinyl sulfone, or a benzoquinone;
• A has the structure of Formula (IIID):
• wherein d and e are integers each independently ranging from 1 to 12;
• Q is a bond, O, S, or N(Rc)(Rd);
• Ra and Rb are independently H, a C1-C4 alkyl group, F, Cl, or N(Rc)(Rd);
• and Re and Rd are independently CH3 or H;
• X is a group comprising between 1 and 30 carbon atoms, wherein the group X comprises a repeat group having Formula (IIIB):
• wherein d and e are integers independently ranging from 1 to 24;
• Q is a bond or O, and wherein the group X a terminates in one of an amine, a carbonyl, an amide, or an imine;
• W is a 5-, 6-membered aromatic or heterocyclic group;
• each R1 is independently selected from the group consisting of H, F, Cl, Br, I, —O-methyl, —O-ethyl, —O-n-propyl, —O-iso-propyl, —O-n-butyl, —O-sec-butyl, and —O-iso-butyl;
• V is a substituted or unsubstituted alkyl group having between 1 and 4 carbon atoms, or a substituted or unsubstituted-O-alkyl group;
• R3 is a phosphate group, an ester group, a sulfate group, a glycoside group, an amide group, or a urea group;
• q is 1;
• s is 0 or an integer ranging from 1 to 4;
• and
• t is 0 or 1,
• wherein when q is 1, the group [V]t-R3 is positioned para relative to the group [A].

A compound having Formula (IE) or (IF):

• wherein
• “Hapten” is digoxigenin, pyrazole, nitropyrazole, benzofuran, oxazole, oxazole sulfonamide, thiazole, or thiazole sulfonamide;
• Y is selected from the group consisting of a carbonyl-reactive group, an amine-reactive group, and a thiol-reactive group, wherein the carbonyl-reactive group is selected from the group consisting of a hydrazine and an amine, wherein the amine-reactive group is a NHS ester, a sulfo-NHS, an isothiocyanate, an isocyanate, an acyl azide, a sulfonyl chloride, an aldehyde, a glyoxal, an epoxide, a oxirane, a carbonate, an aryl halide, an imidoester, or an anhydrides, and wherein the thiol-reactive group is a haloacetyl group, an alkyl halide, a maleimide, an aziridine, an acryloyl, a vinyl sulfone, or a benzoquinone;
• A is a group comprising a branched or unbranched, unsubstituted aliphatic group having between 1 and 4 carbon atoms;
• X is a group comprising between 1 and 30 carbon atoms, wherein the group X comprises a repeat group having Formula (IIIB):
• wherein d and e are integers independently ranging from 1 to 24;
• Q is a bond or O, and wherein the group X a terminates in one of an amine, a carbonyl, an amide, or an imine;
• W is a 6-membered aromatic group or a 5-membered heterocyclic group comprising one or two N, O, or S heteroatoms;
• each R1 is independently selected from the group consisting of H, F, Cl, Br, I, —O-methyl, —O-ethyl, —O-n-propyl, —O-iso-propyl, —O-n-butyl, —O-sec-butyl, and —O-iso-butyl;
• V is a substituted or unsubstituted-O-alkyl group;
• R3 is selected from the group consisting of a phosphate group, an ester group, a sulfate group, a glycoside group, an amide group, a urea group, and a nitro group;
• q 1;
• s is an integer ranging from 1 to 4;
• and
• t is 1,
• wherein when q is 1, the group [V]t-R3 is positioned para relative to the group [A].