Agent for protein precipitation, a method of protein precipitation, a method of protein assay using protein precipitation agent, and a kit for protein assay
AssigneeGeno Technology, Inc.
InventorAftab Alam
Life of this patent
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Abstract
A method of protein precipitation, concentration and removal of non-protein agents from the protein solution wherein the protein solution is treated with a protein-precipitation agent containing an acidic agent, a salt and a precipitate forming agent. After precipitation, the protein precipitate is washed with a water miscible organic solvent agent to remove non-protein agents present in the protein precipitate.
Claims23
A method of preparation of protein sample solution for analysis, wherein the protein sample solution contains one or more of non-protein agents selected from a group consisting of an anionic detergent, a cationic detergent, a non-ionic detergent, a zwitterionic detergent, a sulfobutane, a lipid, a natural product, a salt, and a common laboratory agent, wherein after the preparation of the protein sample the protein in the sample is quantitatively recovered and is substantially free from the non-protein agents originally present in the sample, comprises the following steps:
- (a) treating the protein sample solution with an acidic agent, and one or both agents selected from a group consisting of a salt and a precipitate-forming agent, wherein the treatment of the protein with the precipitate-forming agent follows the treatment of the protein sample solution with the acidic agent;
- (b) centrifuge the protein sample solution of the step (a) at least once to form a tight pellet at the bottom of the tube, remove and discard the supernatant and collect a protein pellet;
- (c) suspend and mix the protein pellet of the step (b) at least once in at least one medium selected from a group consisting of a mixture of aqueous-organic solvent and an organic solvent;
- (d) centrifuge the protein pellet suspension of the step (c) and collect the protein pellet;
- and
- (e) suspend the protein pellet of the step (d) in a protein pellet solubilization reagent buffer, wherein the reagent buffer is provided with an acid neutralizing agent in a sufficient amount to substantially netruralize the acid captured in the protein pellet to facilitate a desired protein solubilization.
A method of preparation of protein sample solution for analysis, wherein the protein sample solution contains one or more of non-protein agents selected from a group consisting of an anionic detergent, a cationic detergents, a non-ionic detergent, a zwitterionic detergent, a sulfobutane, a lipid, a natural product, a salt, and a common laboratory agent, wherein after the preparation of the protein sample the protein in the sample is quantitatively recovered and is substantially free from the non-protein agents originally present in the sample, comprises the following steps:
- (a) treating the protein sample solution with an acidic agent, and one or both agents selected from a group consisting of a salt and a precipitate-forming agent, wherein the treatment of the protein with the precipitate-forming agent follows the treatment of the protein sample solution with the acidic agent;
- (b) centrifuge the protein sample solution of the step (a) to form a tight pellet at the bottom of the tube, remove and discard the supernatant and repeat the centrifugation to remove the residual supernatant with a tipped device and collect a protein pellet;
- (c) suspend and mix the protein pellet of the step (b) at least once in at least one medium selected from a group consisting of a mixture of aqueous-organic solvent and an organic solvent;
- (d) centrifuge the protein pellet suspension of step (c) and collect the protein pellet;
- and
- (e) suspend the protein pellet of the step (d) in a protein pellet solubilization reagent buffer, wherein the reagent buffer is provided with an acid neutralizing agent to neutralize approximately or greater than 0.25nM acid per micro-gram protein in the pellet to facilitate a desired protein solubilization.
A method of total protein assay, wherein the protein sample contains one or more of non-protein agents selected from a group consisting of an anionic detergent, a cationic detergent, a non-ionic detergent, a zerterionic detergent, a sulfobutane, a lipid, a natural product, a salt, and a common laboratory agent, comprises the following steps:
- (a) treating the protein sample solution with an acidic agent, and one or both agents selected from a group consisting of a salt and a precipitate-forming agent, wherein the treatment of the protein with the precipitate-forming agent follows the treatment of the protein sample solution with the acidic agent;
- (b) centrifuge the protein sample solution of the step (a) at least once to form a tight pellet at the bottom of the tube, remove and discard the supernatant and collect a protein pellet;
- (c) suspend the protein pellet of the step (b) with one or more alkaline reagents of a protein assay to produce a characteristic protein reaction;
- and
- (d) compare the color density of the protein color reaction with the color density of a protein reaction of known protein concentration.
Dependencies are inferred from the claim text.