High-performing electrophoresis gels with extended shelf lives
AssigneeBio-Rad Laboratories, Inc.
InventorsShane Petersen, Cory Panattoni, Craig Rowell
Life of this patent
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Abstract
Polyacrylamide gels that offer high resolution as electrophoretic media for protein separations and an improved resistance to hydrolysis upon storage are made by including either taurine, asparagine, or both as an ampholyte, in combination with either tris(hydroxymethyl)-aminomethane or bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane as a buffer, plus other conventional components.
Claims36
A polyacrylamide gel comprising crosslinked polyacrylamide, a buffer selected from the group consisting of tris(hydroxymethyl)aminomethane and bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane, and an ampholyte selected from the group consisting of taurine and asparagine.
A pre-mixed solution for casting to form a polyacrylamide electrophoresis gel, said pre-mixed solution comprising the following ingredients dissolved in water:
- (i) acrylamide monomer,
- (ii) an acrylamide crosslinker,
- (iii) a buffer selected from the group consisting of tris(hydroxymethyl)aminomethane and bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane, and
- (iv) an ampholyte selected from the group consisting of taurine and asparagine.
A method for forming a polyacrylamide electrophoresis gel, said method comprising:
- (a) storing a pre-mixed solution comprising the following ingredients dissolved in water:
- (i) acrylamide monomer,
- (ii) an acrylamide crosslinker,
- (iii) a buffer selected from the group consisting of tris(hydroxymethyl)-aminomethane and bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane, and
- (iv) an ampholyte selected from the group consisting of taurine and asparagine,
- for a period of time exceeding 24 hours;
- (b) adding to said pre-mixed solution a polymerization catalyst to cause said acrylamide and acrylamide crosslinker to polymerize to form a crosslinked polyacrylamide and to thereby transform said pre-mixed solution to a gel.
A method for separating proteins in a liquid sample by electrophoresis, said method comprising:
- (a) loading said sample onto a polyacrylamide gel comprising crosslinked polyacrylamide, a buffer selected from the group consisting of tris(hydroxymethyl)aminomethane and bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane, and an ampholyte selected from the group consisting of taurine and asparagine;
- and
- (b) imposing a voltage differential across said gel to cause said proteins to migrate through said gel at migration rates that vary with the mass, charge, or both of said proteins and to separate into bands within said gel due to said varying migration rates.
Dependencies are inferred from the claim text.