Quantitative peptide or protein assay
AssigneePierce Biotechnology, Inc.
InventorsPaul Jeffrey Haney, Christopher L. Etienne, Sijian Hou, Erum Raja, Ramesh Ganapathy
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Abstract
Peptide and/or protein quantitation methods, kits, and compositions, particularly useful for mass spectrometry, are provided herein based on a bathocuproine-based composition complex such as bathocuproinedisulfonic acid disodium salt hydrate complex. The methods are one-step rapid absorbance methods using small sample volumes. They produce a robust signal with high signal to background ratio and accurately quantitate even complex peptide mixtures with low variability and high sensitivity.
Claims11
A kit for protein or peptide quantitation of a sample comprising:
- (a) a reagent composition comprising sodium carbonate;
- (b) a reagent composition comprising acetonitrile and a complex having the formula:
- wherein
- each of R1 and R2 is independently chosen from C1-C6 straight and branched alkyls, C6-C20 aryl, alkylaryl, and arylalkyls;
- each of R3 and R4 is independently chosen from the group consisting of hydrogen;
- sulfonate salts chosen from sodium, potassium, and lithium;
- phosphonate salts chosen from sodium, potassium, and lithium;
- and carboxylate salts chosen from sodium, potassium, and lithium;
- and
- each of R5 and R6 is
- wherein R7 is independently chosen from the group consisting of hydrogen;
- sulfonate salts chosen from sodium, potassium, and lithium;
- phosphonate salts chosen from sodium, potassium, and lithium;
- and carboxylate salts chosen from sodium, potassium, and lithium;
- with the proviso that at least one of R3, R4, and R7 is not H;
- (c) a reagent composition comprising copper sulfate;
- and
- (d) instructions for the peptide quantitation determination, wherein the determination comprises combining the components (a), (b), and (c) with the sample to form a colored complex, wherein acetonitrile is present in the combination at a concentration ranging from about 10% to about 50%, wherein the kit does not contain an additional reducing agent to reduce excess Cu2+ to Cu1+, and wherein the concentration of peptide or protein in said sample is directly proportional to the amount of colored complex that forms once the sample and components are mixed.
Dependencies are inferred from the claim text.