Hydrolysis-resistant polyacrylamide gels
AssigneeBio-Rad Laboratories, Inc.
InventorsCraig Rowell, Cory Panattoni, Thomas R. Berkelman, Sean Cater, Shane Petersen, Lee Olech, Xuemei Yang
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Abstract
Polyacrylamide gels that offer high resolution in protein separations and are more stable relative to hydrolysis than conventional polyacrylamide gels that rely on Tris or Tris-Bis as buffering agents are made by incorporating triethanolamine in place of most or all of the Tris or Tris-Bis.
Claims20
A polyacrylamide gel comprising:
- crosslinked polyacrylamide;
- triethanolamine;
- a catalytic amount of a polymerization catalyst selected from the group consisting of ammonium persulfate, N,N′-tetramethylenediamine (TEMED), riboflavin, β-dimethylamino-propionitrile, and combinations thereof;
- 0 to 0.3 mM tris(hydroxymethyl)aminomethane;
- and
- 0 to 0.3 mM bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane;
- wherein the polyacrylamide gel has a pH of from about 6.4 to about 7.0, and wherein the polyacrylamide gel is configured to separate and resolve proteins under electrophoretic conditions.
A method for separating a mixture of proteins by electrophoresis, said method comprising:
- (i) loading a gel with a sample of said mixture, wherein said gel comprises:
- crosslinked polyacrylamide;
- triethanolamine;
- a catalytic amount of a polymerization catalyst selected from the group consisting of ammonium persulfate, N,N′-tetramethylenediamine (TEMED), riboflavin, β-dimethylamino-propionitrile, and combinations thereof;
- 0 to 0.3 mM tris(hydroxymethyl)aminomethane;
- and
- 0 to 0.3 mM bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane;
- wherein the gel has a pH of from about 6.4 to about 7.0;
- and
- (ii) imposing a voltage across said gel to cause said proteins to migrate through said gel at differential rates until separated into bands within said gel.
A reaction mixture for casting a polyacrylamide gel configured to separate and resolve proteins by electrophoresis, said reaction mixture comprising:
- crosslinked polyacrylamide;
- triethanolamine;
- a catalytic amount of a polymerization catalyst selected from the group consisting of ammonium persulfate, N,N′-tetramethylenediamine (TEMED), riboflavin, β-dimethylamino-propionitrile, and combinations thereof;
- 0 to 0.3 mM tris(hydroxymethyl)aminomethane;
- and
- 0 to 0.3 mM bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane.
Dependencies are inferred from the claim text.