Reagents and procedures for multi-label high-specificity labeling

A method for labeling and/or detecting a target material comprising:

• (a) providing:
• at least one target material;
• a first peptide tag bound to said at least one target material;
• and a second peptide tag bound to said at least one target material;
• (b) reacting said at least one target material with the following under suitable conditions to allow labeling of the target material:
• (i) a first conjugate having a detectable group and two pendant phenylarsine moieties comprising a first tag binding group;
• wherein said first conjugate preferentially associates with said first peptide tag;
• and
• (ii) a second conjugate having a detectable group and two pendant phenylarsine moieties comprising a second tag binding group;
• wherein said second conjugate preferentially associates with said second peptide tag;
• wherein the mean distance and/or mean angle between said pendant phenylarsine moieties in said first conjugate is different from the mean distance and/or mean angle between said pendant phenylarsine moieties in said second conjugate.

A method for monitoring a binding process comprising:

• (a) reacting a first component of a specific binding pair with a second component of said pair;
• said pair having bound thereto a first peptide tag and a second peptide tag and being labeled with the following:
• (i) a first conjugate having a detectable group and two pendant phenylarsine moieties comprising a first tag binding group;
• wherein said first conjugate preferentially associates with said first peptide tag;
• and
• (ii) a second conjugate having a detectable group and two pendant phenylarsine moieties comprising a second tag binding group;
• wherein said second conjugate preferentially associates with said second peptide tag;
• wherein the mean distance and/or mean angle between said pendant phenylarsine moieties in said first conjugate is different from the mean distance and/or mean angle between said pendant phenylarsine moieties in said second conjugate;
• and
• (b) monitoring for a change in a property of said detectable group on at least one of said first and second conjugates in the presence of both components of said pair, as compared to in the presence of only one of said components of said pair.

A method for determining the localization, concentration or interactions of at least one target material of interest on or within a cell, tissue, organ or organism, said method comprising:

• (a) providing a cell, tissue, organ or organism containing:
• at least one target material;
• a first peptide tag bound to said at least one target material;
• and a second peptide tag bound to said at least one target material;
• (b) contacting said cell, tissue, organ, or organism with the following under suitable conditions to allow labeling of said at least one target material:
• a first conjugate having a detectable group and two pendant phenylarsine moieties comprising a first tag binding group;
• wherein said first conjugate preferentially associates with said first peptide tag;
• and
• a second conjugate having a detectable group and two pendant phenylarsine moieties comprising a second tag binding group;
• wherein said second conjugate preferentially associates with said second peptide tag;
• wherein the mean distance and/or mean angle between said pendant phenylarsine moieties in said first conjugate is different from the mean distance and/or mean angle between said pendant phenylarsine moieties in said second conjugate;
• and
• (c) detecting said label, thereby determining the localization, concentration, or interactions of said target material.

An assay method for monitoring a reaction, comprising:

• (a) reacting a first reactant of a reactant pair with a second reactant of said pair;
• said reactant pair having bound thereto a first peptide tag and a second peptide tag and being labeled with the following:
• (i) a first conjugate having a detectable group and two pendant phenylarsine moieties comprising a first tag binding group;
• wherein said first conjugate preferentially associates with said first peptide tag;
• and
• (ii) a second conjugate having a detectable group and two pendant phenylarsine moieties comprising a second tag binding group;
• wherein said second conjugate preferentially associates with said second peptide tag;
• wherein the mean distance and/or mean angle between said pendant phenylarsine moieties in said first conjugate is different from the mean distance and/or mean angle between said pendant phenylarsine moieties in said second conjugate;
• and
• (b) monitoring for a change in a property of said detectable group on at least one of said first and second conjugates in the presence of both of said reactants, as compared to in the presence of only one of said reactants.

A method for isolating at least one target material of interest comprising:

• (a) providing a solution including:
• at least one target material;
• a first peptide tag bound to said at least one target material;
• and a second peptide tag bound to said at least one target material;
• (b) contacting said provided solution with first and second conjugates immobilized on a solid support under suitable conditions to allow binding of said at least one target material to said immobilized conjugates;
• said first conjugate having a detectable group and two pendant phenylarsine moieties comprising a first tag binding group;
• wherein said first conjugate preferentially associates with said first peptide tag;
• said second conjugate having a detectable group and two pendant phenylarsine moieties comprising a second tag binding group;
• wherein said second conjugate preferentially associates with said second peptide tag;
• wherein the mean distance and/or mean angle between said pendant phenylarsine moieties in said first conjugate is different from the mean distance and/or mean angle between said pendant phenylarsine moieties in said second conjugate;
• and
• (c) eluting said target material with a low-molecular weight monothiol or low-molecular weight dithiol.

A kit comprising:

• (a) at least one vector including sequence encoding a first peptide tag and/or a second peptide tag;
• (b) a first conjugate having a detectable group and two pendant phenylarsine moieties comprising a first tag binding group;
• wherein said first conjugate preferentially associates with said first peptide tag;
• (c) a second conjugate having a detectable group and two pendant phenylarsine moieties comprising a second tag binding group;
• wherein said second conjugate preferentially associates with said second peptide tag;
• wherein the mean distance and/or mean angle between said pendant phenylarsine moieties in said first conjugate is different from the mean distance and/or mean angle between said pendant phenylarsine moieties in said second conjugate.

A composition comprising:

• (i) at least one target material;
• (ii) a first peptide tag bound to said at least one target material and being of the form:
• C(X)iC(X)jC(X)kC, wherein C is cysteine, X is any amino acid, and i, j, and k are each independently selected from 0 or an integer from 1-8;
• (iii) a second peptide tag bound to said at least one target material and being of the form C(X)iC, wherein X is any amino acid and i is 0-6.

A composition comprising at least one isolated or recombinant nucleic acid encoding:

• (i) at least one target material;
• (ii) a first peptide tag bound to the at least one target material and being of the form:
• C(X)iC(X)jC(X)kC, wherein C is cysteine, X is any amino acid, and i, j, and k are each independently selected from 0 or an integer from 1-8;
• and (iii) a second peptide tag bound to the at least one target material and being of the form C(X)iC, wherein X is any amino acid and i is 0-6.