Electrophoresis buffer for faster migration, improved resolution and extended shelf-life

Abstract

There is provided an electrolyte solution for extending shelf life, and/or accelerating or improving resolution or improving transfer efficacy for blot applications, or accelerating and improving resolution, or accelerating and improving transfer efficacy of gel electrophoresis and containing Tris(hydroxymethyl)aminomethane (TRIS), at least one zwitterion, and water. The electrolyte solution may be used in buffer systems for gel electrophoresis and the preparation of gels for gel electrophoresis, such as Western blot.

1 .- 43 . (canceled) 
     
     
         44 . An electrolyte solution comprising:
 Tris(hydroxymethyl)aminomethane (TRIS);   at least one zwitterion chosen from 2-amino-2methyl-1,3-propanediol (AMPD), N,N-bis[2-hydroxyethyl]-2-aminoethanesulphonic acid (BES), N-Glycylglycine (Gly-Gly), 4-(2-hydroxyethyl)piperazine-1-propanesulfonic acid (EPPS or HEPPS), and 3-(cyclohexylamino)-2-hydroxy-1-propanesulfonic acid (CAPSO), N-(1,1-Dimethyl-2-hydroxyethyl)-3-amino-2-hydroxypropanesulfonic acid (AMPSO), 3-(cyclohexylamino)-1-propanesulfonic acid (CAPS), 2-(cyclohexylamino)ethanesulfonic acid (CHES), 3-[N-Bis(hydroxyethyl)amino]-2-hydroxypropanesulfonic acid (DIPSO), N-(2-Hydroxyethyl)piperazine-N′-(4-butanesulfonic acid) (HEPBS), 4-(N-Morpholino)butanesulfonic acid (MOBS), piperazie-N,N′-bis[2-hydroxypropanesulphonic]acid (POPSO), N-Tris(hydroxymethyl)methyl-4-aminobutanesulfonic acid (TABS), N-Tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid (TAPS), (2-[2-hydroxy-1,1-bis(hydroxymethyl)ethylamino]ethanesulphonic acid) (TES), (2-[2-acetamino]-2-aminoethanesulphonic acid) (ACES), (1,4-piperazinediethanesulphonic acid) (PIPES), and Piperazine-N,N′-bis(3-propanesulfonic Acid) (PIPPS);   water, and   a chelating agent having the name: ethylenediaminetetraacetate (EDTA), ethylene glycol tetraacetic acid (EGTA), trisodium nitrilotriacetate, hydroxyethyl ethylenediamine trisodium acetate (trisodium HEDTA), diethylene triamino pentasodium acetate or uramil disodium acetate, wherein said EDTA is at a concentration of 0.5% (w/v) or less;   wherein the pH of the electrolyte solution is from about 8.0 to about 8.8,   wherein said electrolyte solution is for at least one of:   a shorter electrophoresis time of a gel electrophoresis,   an improved resolution of a gel electrophoresis,   an improved transfer efficacy of a gel electrophoresis, and   a shorter transfer time of a gel electrophoresis,   compared to a Tris-Glycine-SDS electrolyte solution at pH 8.3.   
     
     
         45 . The electrolyte solution of  claim 44 , further comprising methanol. 
     
     
         46 . The electrolyte solution of  claim 45 , wherein said methanol is up to 20%. 
     
     
         47 . The electrolyte solution of  claim 44 , further comprising sodium dodecyl sulphate (SDS). 
     
     
         48 . The electrolyte solution of  claim 47 , wherein sodium dodecyl sulphate (SDS) is 0.5% (wt/vol) or less. 
     
     
         49 . The electrolyte solution of  claim 48 , wherein the concentration of sodium dodecyl sulphate (SDS) is 0.1% (wt/vol) or less. 
     
     
         50 . The electrolyte solution of  claim 44 , wherein a concentration of said chelating agent is 0.5% (wt/vol) or less. 
     
     
         51 . The electrolyte solution of  claim 50 , wherein a concentration of said chelating agent is 0.05% (wt/vol) or less. 
     
     
         52 . The electrolyte solution of  claim 51 , wherein a concentration of said chelating agent is 0.03%. 
     
     
         53 . The electrolyte solution of  claim 44 , wherein the concentration of Tris(hydroxymethyl)aminomethane (TRIS) is from about 10 mM to about 500 mM. 
     
     
         54 . The electrolyte solution of  claim 44 , wherein the concentration of the zwitterion is from about 1 mM to about 500 mM. 
     
     
         55 . The electrolyte solution of  claim 44 , wherein the concentration of Tris(hydroxymethyl)aminomethane (TRIS) is from about 50 mM to about 150 mM. 
     
     
         56 . The electrolyte solution of  claim 44 , wherein the concentration of Tris(hydroxymethyl)aminomethane (TRIS) is from about 50 mM to about 250 mM. 
     
     
         57 . The electrolyte solution of  claim 44 , wherein the concentration of Tris(hydroxymethyl)aminomethane (TRIS) is 150 mM. 
     
     
         58 . The electrolyte solution according to  claim 44 , wherein said zwitterions is a combination of 4-(2-hydroxyethyl)piperazine-1-propanesulfonic acid (EPPS or HEPPS), (2-[2-acetamino]-2-aminoethanesulphonic acid) (ACES), and (1,4-piperazinediethanesulphonic acid) (PIPES). 
     
     
         59 . An electrophoresis gel comprising the electrolyte solution of  claim 44 , wherein the gel is chosen from an acrylamide gel at a concentration from less than 1% (wt/vol) to about 25% (wt/vol), an agarose gel at a concentration from about 0.5% (wt/vol) to about 3% (wt/vol) and a combination thereof. 
     
     
         60 . The electrophoresis gel of  claim 59 , further comprising sodium dodecyl sulphate (SDS). 
     
     
         61 . The electrophoresis gel of  claim 59 , wherein the pH is from about 8.0 to about 8.8. 
     
     
         62 . A method of accelerating or improving the resolution, or improving transfer efficacy, or accelerating and improving the resolution, or accelerating and improving transfer efficacy of the electrophoretic separation of at least one sample comprising the step of:
 applying a voltage to an electrolyte solution according to  claim 44 , in contact with an electrophoresis gel containing the at least one sample therein.