Bis-biotinylation tags

Abstract

Multi-biotinylated reactants are provided which can be used in divalent complexes for various applications such as colocalization, labeling, immobilization, and purification. Methods for constructing, purifying, and using the bis-biotinylated reactants are also provided. In certain embodiments, two bis-biotinylated reactants are bound to a single streptavidin tetramer to provide a complex having a 1:1 stoichiometry with respect to the bis-biotinylated reactants.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of US Provisional Application No. 61 / 835,311, filed June 14, 2013, which is hereby incorporated by reference in its entirety for all purposes. [0003] Statement Regarding Federally Funded Research [0004] Not applied. Background technique [0005] A common task in molecular biology is to identify and quantify the presence of proteins in complex mixtures. For example, to identify the expression level of a protein of interest, a Western blot can be performed in which protein extracts are run on a gel and stained with antibodies directed against defined epitopes of the protein of interest. A defined epitope may be a specific sequence or structure found in a native protein, or it may be a tag introduced during cloning, such as a "FLAG tag", against which antibodies are commercially available. A peroxidase-conjugated secondary antibody specific for the primary antibody bound to the pro...

AI Technical Summary

Problems solved by technology

However, Howarth's method involves mixing 2 recombinant streptavidins and is tedious

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