# US 8,785,203 B2 · Protein detection reagents and methods with dyes and dextrins > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,785,203 B2 - Document ID: US8785203B2 - Status: Granted - Kind: B2 - Application number: 13/278,699 - Filed: 2011-10-21 - Published: 2012-07-26 - Granted: 2014-07-22 - Earliest claimed: 2006-04-28 - Assignees: Expedeon Limited - Applicants: Daniel Brian Jones; Heikki Lanckriet - Inventors: Daniel Brian Jones; Heikki Lanckriet - Web page: https://patentgrep.com/patent/US8785203B2 ## CPC - G01N 33/6833: {Copper, e.g. Folin-, Lowry-, biuret methods} - G01N 33/6839: {involving dyes, e.g. Coomassie blue, bromcresol green} - Y10T 436/25125: Digestion or removing interfering materials - 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) - Y10T: TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION ## Abstract The invention provides reagents, methods and kits for detection of proteins and quantitative determination of protein concentration. The reagents comprise a protein-complexing dye, such as a Coomassie dye and one or more dextrins, for the elimination of interference caused by detergents. ## Claims ### Claim 1 A method of detecting and/or quantifying protein comprising contacting a protein-containing sample with a solution comprising: (a) a protein complexing dye; and (b) one dextrin selected from α-cyclodextrins, γ-cyclodextrins, linear dextrins and derivatives thereof, or two or more dextrins; and detecting and/or quantifying dye/protein complex formation, wherein the protein is in the presence of a detergent. ### Claim 2 (depends on 1) The method according to claim 1, wherein the protein is in solution or the protein is provided on a support. ### Claim 3 (depends on 2) The method according to claim 2, wherein the support is a gel, sol, chromatography plate, filter paper, nitrocellulose membrane or resin. ### Claim 4 (depends on 3) The method according to claim 3, wherein the support is a polyacrylamide gel or agarose gel and the protein has been subjected to separation using an electric field. ### Claim 5 (depends on 2) The method according to claim 2, wherein the support comprises a detergent and/or wherein contacting is performed in the presence of a detergent. ### Claim 6 (depends on 1) The method according to claim 1, wherein detecting and/or quantifying comprises detecting a change in absorption or emission spectra of the dye/protein complex. ### Claim 7 (depends on 6) The method according to claim 6, wherein detecting and/or quantifying comprises detecting a colour change. ### Claim 8 (depends on 6) The method according to claim 6, wherein the protein-complexing dye is a Coomassie G-250 dye and absorbance is measured at a wavelength of about 595 nm. ### Claim 9 (depends on 1) The method according to claim 1, wherein quantifying is performed by measuring absorbance over time. ### Claim 10 (depends on 6) The method according to claim 6, wherein the absorbance or emission measured is compared with a standard value, standard set of values, or standard curve. ### Claim 11 (depends on 1) The method according to claim 1, further comprising providing a support comprising protein, prior to the contacting step. ### Claim 12 A method comprising contacting a protein-containing sample in the presence of a detergent with a protein binding dye, and one dextrin selected from α-cyclodextrins, γ-cyclodextrins, linear dextrins and derivatives thereof; and detecting and/or quantifying dye/protein complex formation, wherein the protein is in the presence of a detergent. ### Claim 13 A method of detecting and/or quantifying protein comprising contacting a protein-containing sample in the presence of a detergent with a protein binding dye, and two or more dextrins; and detecting and/or quantifying dye/protein complex. ### Claim 14 (depends on 1) The method according to claim 1, wherein the one dextrin or two or more dextrins are provided as a reagent comprising a protein-complexing dye and one dextrin. ### Claim 15 (depends on 14) The method according to claim 14, wherein the reagent is a reagent concentrate. ### Claim 16 (depends on 14) The method according to claim 14, wherein the reagent further comprises an acid with a pKa of 4 or less. ### Claim 17 (depends on 14) The method according to claim 14, wherein the reagent further comprises a solubilizing agent. ### Claim 18 (depends on 14) The method according to claim 14, wherein the reagent does not comprise a protein. ### Claim 19 (depends on 14) The method according to claim 14, wherein the dye is a Coomassie dye. ### Claim 20 (depends on 1) The method according to claim 1, wherein the one dextrin, or two or more dextrins concentration is in the range of from about 0.01 to about 200 mg/ml. ### Claim 21 (depends on 14) The method according to claim 14, wherein the reagent further comprises one or more alcohols, and wherein the alcohol concentration is from about 0.1% to about 10% v/v. ### Claim 22 (depends on 13) The method according to claim 13, wherein the mixture of two or more dextrins is selected from the group consisting of α-cyclodextrins, β-cyclodextrins, γ-cyclodextrins, cycloamylose, linear dextrins and derivatives thereof. ### Claim 23 (depends on 14) The method according to claim 14, wherein the reagent is provided as a multipart system. ### Claim 24 A method of detecting and/or quantifying protein comprising contacting a protein-containing sample with a solution comprising: (a) a protein-complexing dye, wherein the dye is a Coomassie dye; and two or more dextrins; and detecting and/or quantifying dye/protein complex formation, wherein the protein is on a support. ### Claim 25 (depends on 24) A method according to claim 24, wherein the dye is a Coomassie dye. ### Claim 26 (depends on 24) A method according to claim 24, wherein the support is a gel, sol, chromatography plate, filter paper, nitrocellulose membrane or resin. ### Claim 27 (depends on 26) The method according to claim 26, wherein the support is a polyacrylamide gel or agarose gel and the protein has been subjected to separation using an electric field. ### Claim 28 (depends on 24) The method according to claim 24, wherein the support comprises a detergent and/or wherein contacting is performed in the presence of a detergent. ### Claim 29 A method of detecting and/or quantifying protein comprising contacting a protein-containing sample with a solution comprising: (a) a protein-complexing dye; and (b) one dextrin selected from α-cyclodextrins, γ-cyclodextrins, linear dextrins and derivatives thereof; and detecting and/or quantifying dye/protein complex formation, wherein the protein is on a support. ### Claim 30 (depends on 29) A method according to claim 29, wherein the dye is a Coomassie dye. ### Claim 31 (depends on 30) The method according to claim 30, wherein the Coomassie dye is a Coomassie G-250 dye. ### Claim 32 (depends on 29) A method according to claim 29, wherein the support is a gel, sol, chromatography plate, filter paper, nitrocellulose membrane or resin. ### Claim 33 (depends on 32) The method according to claim 32, wherein the support is a polyacrylamide gel or agarose gel and the protein has been subjected to separation using an electric field. ### Claim 34 (depends on 29) The method according to claim 29, wherein the support comprises a detergent and/or wherein contacting is performed in the presence of a detergent.