One-step reduction and alkylation of proteins
AssigneeIndiana Proteomics Consortium, LLC
InventorsJohn Hale, Michael Knierman
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Abstract
The present invention relates generally to the field of analyzing proteins and more specifically to the reduction and alkylation of protein samples during protein analysis. A novel composition and method for reducing and alkylating proteins is disclosed. A novel reagent including a combination of a volatile reducing agent, a volatile alkylating agent, and a volatile solvent is used for a one-step reduction and alkylation of proteins allowing the protein sample to remain in the same container during the reduction and alkylation processes.
Claims54
A composition configured to modify proteins having disulfide bonds, the composition comprising:
- a single volatile reagent for breaking the disulfide bonds and preventing reformation thereof.
A method of modifying a protein comprising the steps of:
- selecting a protein;
- providing a volatile reagent to reduce and alkylate the protein, the reagent including a sufficient amount of solvent to substantially dissolve the protein;
- and
- dissolving the protein in the volatile reagent to produce a first mixture.
The method of 23, wherein the chelated metal is copper citrate.
A kit for one-step reduction and alkylation of a protein sample, the kit including:
- a substantially volatile reducing agent to reduce the protein sample;
- a substantially volatile alkylating agent to alkylate the protein sample;
- and
- a substantially volatile solvent to substantially dissolve the protein sample.
A composition for one-step reduction and alkylation of a protein sample, the composition including:
- a sufficiently volatile reducing agent to sufficiently reduce the protein sample;
- a sufficiently volatile alkylating agent to sufficiently alkylate the protein sample;
- and
- a sufficiently volatile solvent to sufficiently dissolve the protein sample.
A method for producing a volatile reagent for modifying a protein sample, the method including the steps of:
- selecting a substantially volatile reducing agent for sufficiently reducing the protein sample;
- selecting a substantially volatile alkylating agent for sufficiently alkylating the protein sample;
- selecting a substantially volatile solvent for sufficiently dissolving the protein sample;
- mixing the reducing agent, alkylating agent, and solvent to form a mixture.
A method of a modifying protein comprising the steps of:
- selecting a protein;
- providing a substantially volatile reducing agent for reducing the protein;
- providing a substantially volatile alkylating agent for alkylating the protein;
- and
- providing a substantially volatile solvent for dissolving the protein.
Dependencies are inferred from the claim text.