Native protein electrophoresis and methods of use
A method of characterizing proteins has been developed that includes providing a sample that contains a plurality of proteins to be characterized, wherein at least a first protein of the plurality of proteins is in its native form. Additionally, the method includes contacting the sample containing the plurality of proteins with a solution to form a sample solution, and then contacting the sample solution with a gel. The plurality of proteins is subsequently separated via electrophoresis within the gel, which includes an electrophoresis solution.
1. A method of characterizing proteins, the method comprising:
(a) providing a sample comprising a plurality of proteins to be characterized, wherein the plurality of proteins are in their native form;
(b) contacting the sample from step (a) with a solution to form a sample solution;
(c) contacting the sample solution of step (b) with a gel; and
(d) separating the plurality of proteins within the sample solution by electrophoresis within the gel, wherein the gel comprises an electrophoresis solution,
wherein the electrophoresis solution of step (d) comprises a detergent at a concentration of 0.03 wt % to 0.04 wt %, and the plurality of proteins maintain their native form in steps (a)-(d).
2. The method of claim 1 , wherein the sample solution of step (b) is substantially free of detergent.
3. The method of claim 1 , wherein the sample solution of step (b) is at a temperature of less than 30° C.
4. The method of claim 1 , wherein the sample solution of step (b) is at a temperature of less than 10° C.
5. The method of claim 1 , wherein the electrophoresis solution of step (d) comprises the detergent at a concentration of 0.0375 wt %.
6. The method of claim 1 , wherein the detergent comprises an alkyl-sulfate compound.
7. The method of claim 6 , wherein the alkyl-sulfate compound comprises sodium dodecyl sulfate.
8. The method of claim 1 , wherein separating the plurality of proteins as in step (d) is performed at 4° C.
9. The method of claim 1 , wherein separating the plurality of proteins within the sample solution by electrophoresis within the gel comprises separating the plurality of proteins within the sample solution by electrophoresis within the gel such that two proteins which differ in molecular weight by 2 kDa can be distinguished from one another on a 12% polyacrylamide gel.
10. The method of claim 1 , wherein the native form comprises enzymatic activity.
11. The method of claim 10 , wherein the enzymatic activity is provided at least in part by an enzyme that is chosen from the group consisting of dehydrogenase, galactosidase, dismutase, phosphatase, urease, oxidase, anhydrase, kinase, protease, and synthase.
12. The method of claim 1 , wherein the plurality of proteins maintain their native interactions with at least one metal ion, wherein the at least one metal ion is an ion of a metal chosen from the group consisting of copper, magnesium, manganese, molybdenum, nickel, and zinc.