Electrophoresis buffer for extending the useful electrophoresis life of an electrophoresis gel

Abstract

There is provided an electrolyte solution for extending useful electrophoresis life of an electrophoresis gel containing Tris(hydroxymethyl)aminomethane (TRIS), at least one zwitterion, and water. The electrolyte solution may be used in buffer systems for gel electrophoresis, such as SDS-PAGE.

1 .- 28 . (canceled) 
     
     
         29 . An electrolyte solution for extending a useful electrophoresis life of an electrophoresis gel comprising:
 at least one zwitterion chosen from 2-amino-2methyl-1,3-propanediol (AMPD), N-(1,1-Dimethyl-2-hydroxyethyl)-3-amino-2-hydroxypropanesulfonic acid (AMPSO), N-Glycylglycine (Gly-Gly), 4-(2-hydroxyethyl)piperazine-1-propanesulfonic acid (EPPS or HEPPS), 3-(cyclohexylamino)-1-propanesulfonic acid (CAPS), 3-(cyclohexylamino)-2-hydroxy-1-propanesulfonic acid (CAPSO), 2-(cyclohexylamino)ethanesulfonic acid (CHES), N,N-bis[2-hydroxyethyl]-2-aminoethanesulphonic acid (BES), (2-[2-hydroxy-1,1-bis(hydroxymethyl)ethylamino] ethanesulphonic acid) (TES), N-Tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid (TAPS), 3-N-Morpholino propanesulfonic acid (MOPS), and Piperazine-N, N′-bis(2-hydroxypropanesulfonic acid) (POPSO);   water; and   1.0% (wt/vol) or less of an anionic surfactant selected from the group consisting of sodium dodecyl sulphate (SDS), lithium dodecyl sulphate (LDS), or combinations thereof,   wherein the pH of the electrolyte solution is from about 8.0 to about 11.0.   
     
     
         30 . The electrolyte solution of  claim 29 , wherein the zwitterion is 4-(2-hydroxyethyl)piperazine-1-propanesulfonic acid (EPPS or HEPPS). 
     
     
         31 . The electrolyte solution of  claim 29 , wherein the zwitterion is chosen from N-Glycylglycine (Gly-Gly) and 3-(cyclohexylamino)-2-hydroxy-1-propanesulfonic acid (CAPSO). 
     
     
         32 . The electrolyte solution of  claim 29 , further comprising Tris(hydroxymethyl)aminomethane (TRIS). 
     
     
         33 . The electrolyte solution of  claim 29 , wherein the concentration of the zwitterion is from about 1 mM to about 500 mM. 
     
     
         34 . The electrolyte solution of  claim 33 , wherein the concentration of the zwitterion is from about 10 mM to about 500 mM. 
     
     
         35 . The electrolyte solution of  claim 33 , wherein the concentration of the zwitterion is from about 25 mM to about 75 mM. 
     
     
         36 . The electrolyte solution of  claim 33 , wherein the concentration of the zwitterion is from about 50 mM to about 100 mM. 
     
     
         37 . The electrolyte solution of  claim 36 , wherein the concentration of the zwitterion is 50 mM. 
     
     
         38 . The electrolyte solution of  claim 36 , wherein the concentration of the zwitterion is 100 mM. 
     
     
         39 . The electrolyte solution of  claim 29 , wherein the concentration of anionic surfactant is 0.5% (wt/vol) or less. 
     
     
         40 . The electrolyte solution of  claim 39 , wherein the concentration of anionic surfactant is 0.1% (wt/vol) or less. 
     
     
         41 . The electrolyte solution of  claim 39 , wherein the concentration of anionic surfactant is 0.1% (wt/vol). 
     
     
         42 . The electrolyte solution of  claim 29 , further comprising a chelating agent selected from the group consisting of ethylene diamine tetraacetate (EDTA), ethylene glycol tetraacetic acid (EGTA), trisodium nitrilotriacetate, hydroxyethyl ethylenediamine trisodium acetate (trisodium HEDTA), diethylene triamino pentasodium acetate, and uramil disodium acetate. 
     
     
         43 . The electrolyte solution of  claim 42 , wherein the concentration of said chelating agent is from about 0.03% to about 0.5% (wt/vol). 
     
     
         43 . The electrolyte solution of  claim 43 , wherein the concentration of said chelating agent is from about 0.05% to about 0.4% (wt/vol). 
     
     
         44 . The electrolyte solution of  claim 29 , further comprising Tris(hydroxymethyl)aminomethane (TRIS). 
     
     
         45 . The electrolyte solution of  claim 44 , wherein the concentration of Tris(hydroxymethyl)aminomethane (TRIS) is from about 10 mM to about 500 mM. 
     
     
         46 . The electrolyte solution of  claim 45 , wherein the concentration of Tris(hydroxymethyl)aminomethane (TRIS) is from about 10 mM to about 150 mM. 
     
     
         47 . The electrolyte solution of  claim 45 , wherein the concentration of Tris(hydroxymethyl)aminomethane (TRIS) is 150 mM. 
     
     
         48 . A method of extending a useful electrophoresis life of an electrophoresis gel during an electrophoretic separation of at least one sample comprising the step of:
 applying a voltage to an electrolyte solution according to  claim 29 , in contact with an electrophoresis gel containing the at least one sample therein.   
     
     
         49 . A method of extending a useful electrophoresis life of an electrophoresis gel during an electrophoretic separation of at least one sample comprising the step of:
 adding to an electrophoresis buffer containing an electrolyte solution according to  claim 29  at least one different zwitterion chosen from 2-amino-2methyl-1,3-propanediol (AMPD), N-(1,1-Dimethyl-2-hydroxyethyl)-3-amino-2-hydroxypropanesulfonic acid (AMPSO), N-Glycylglycine (Gly-Gly), 4-(2-hydroxyethyl)piperazine-1-propanesulfonic acid (EPPS or HEPPS), 3-(cyclohexylamino)-1-propanesulfonic acid (CAPS), 3-(cyclohexylamino)-2-hydroxy-1-propanesulfonic acid (CAPSO), 2-(cyclohexylamino)ethanesulfonic acid (CHES), N,N-bis[2-hydroxyethyl]-2-aminoethanesulphonic acid (BES), (2-[2-hydroxy-1,1-bis(hydroxymethyl)ethylamino] ethanesulphonic acid) (TES), N-Tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid (TAPS) and 3-N-Morpholino propanesulfonic acid (MOPS).   
     
     
         50 . The method according to  claim 49 , wherein said zwitterion is 4-(2-hydroxyethyl)piperazine-1-propanesulfonic acid (EPPS or HEPPS). 
     
     
         51 . The method according to  claim 49 , wherein the zwitterion is chosen from N-Glycylglycine (Gly-Gly) and 3-(cyclohexylamino)-2-hydroxy-1-propanesulfonic acid (CAPSO). 
     
     
         52 . The method according to  claim 49 , further comprising the step of applying a voltage to said electrophoresis buffer in contact with an electrophoresis gel containing the at least one sample therein.