# US 10,119,934 B2 · Hydrolysis-resistant polyacrylamide gels > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,119,934 B2 - Document ID: US10119934B2 - Status: Granted - Kind: B2 - Application number: 15/272,257 - Filed: 2016-09-21 - Published: 2017-03-09 - Granted: 2018-11-06 - Earliest claimed: 2008-09-02 - Assignees: Bio-Rad Laboratories, Inc. - Applicants: Bio-Rad Laboratories, Inc. - Inventors: Craig Rowell; Cory Panattoni; Thomas R. Berkelman; Sean Cater; Shane Petersen; Lee Olech; Xuemei Yang - Web page: https://patentgrep.com/patent/US10119934B2 ## CPC - G01N 27/44747: {Composition of gel or of carrier mixture} - B01D 57/02: by electrophoresis (treatment of water, waste water, sewage or sludge by electrophoresis C02F1/469; electrophoretic production of compounds or non-metals C25B7/00; investigating or analysing materials by using electrophoresis G01N27/26) - C08F 220/56: Acrylamide; Methacrylamide - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - B01D: SEPARATION (separating solids from solids by wet methods B03B, B03D; by pneumatic jigs or tables B03B; by other dry methods B07; magnetic or electrostatic separation of solid materials from solid materials or fluids, separation by high-voltage electric fields B03C; centrifuges, vortex apparatus B04; presses per se for squeezing-out liquid from liquid-containing material B30B9/02; treatment of water C02F, e.g. softening by ion-exchange C02F1/42; {arrangements of air intake cleaners in gas turbine plants F02C7/05} ; arrangements or mounting of filters in air-conditioning, air-humidification or ventilation F24F13/28) - C08F: MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS ## 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. ## Claims ### Claim 1 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. ### Claim 2 (depends on 1) The polyacrylamide gel of claim 1 wherein said triethanolamine constitutes from about 50 mM to about 150 mM of said gel. ### Claim 3 (depends on 1) The polyacrylamide gel of claim 1 wherein said gel is devoid of tris(hydroxymethyl)aminomethane and devoid of bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane. ### Claim 4 (depends on 1) The polyacrylamide gel of claim 1 further comprising an ampholyte. ### Claim 5 (depends on 4) The polyacrylamide gel of claim 4 wherein said ampholyte is glycine at a concentration of from about 50 mM to about 500 mM. ### Claim 6 (depends on 5) The polyacrylamide gel of claim 5 further comprising a conjugate ampholyte at a proportion of from about 0.1 mole percent to about 65 mole percent relative to said glycine. ### Claim 7 (depends on 6) The polyacrylamide gel of claim 6 wherein said conjugate ampholyte is taurine and is at a proportion of from about 20 mole percent to about 60 mole percent relative to said glycine. ### Claim 8 (depends on 5) The polyacrylamide gel of claim 5 further comprising a weak acid at a concentration of from about 10 mM to about 500 mM. ### Claim 9 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. ### Claim 10 (depends on 9) The method of claim 9 wherein said triethanolamine constitutes from about 50 mM to about 250 mM of said gel. ### Claim 11 (depends on 9) The method of claim 9 wherein said gel is devoid of tris(hydroxymethyl)aminomethane and devoid of bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane. ### Claim 12 (depends on 9) The method of claim 9 wherein said gel further comprises glycine at a concentration of from about 50 mM to about 500 mM. ### Claim 13 (depends on 12) The method of claim 12 wherein said gel further comprises taurine at a proportion of from about 20 mole percent to about 60 mole percent relative to said glycine. ### Claim 14 (depends on 9) The method of claim 9 wherein said triethanolamine constitutes from about 50 mM to about 150 mM of said gel, and said gel further comprises: glycine at a concentration of from about 50 mM to about 500 mM; taurine at a proportion of from about 20 mole percent to about 60 mole percent relative to said glycine; and a weak acid at a concentration of from about 10 mM to about 500 mM. ### Claim 15 (depends on 9) The method of claim 9 wherein said voltage is from about 50 V/cm to about 100 V/cm. ### Claim 16 (depends on 1) The polyacrylamide gel of claim 1 wherein the gel resides in a running buffer comprising tris(hydroxymethyl)aminomethane and glycine. ### Claim 17 (depends on 1) The polyacrylamide gel of claim 1 wherein the gel resides in a running buffer that is substantially free of triethanolamine. ### Claim 18 (depends on 4) The method of claim 4, further comprising: immersing at least a portion of the gel in a running buffer comprising tris(hydroxymethyl)aminomethane and glycine. ### Claim 19 (depends on 4) The method of claim 4, further comprising: immersing at least a portion of the gel in a running buffer that does not comprise triethanolamine. ### Claim 20 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.