Protein staining compositions and methods
AssigneePromega Corporation
InventorsJohn W. Shultz, David L. Leland
Life of this patent
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Abstract
The present invention provides novel dyes for staining proteins, particularly those in electrophoretic gels. The dyes of the invention are derivatives of Coomassie.RTM. dyes. The invention also provides quantitative and qualitative methods for detecting proteins by staining the proteins with a dye of the invention.
Claims12
A method of detecting a protein in a polyacrylamide or agarose support, in which the protein has been electrophoresed, comprising staining the protein in the support with a dye of Formula X ##STR3## wherein R.sup.10, R.sup.20, and R.sup.30 are independently selected from the group consisting of hydrogen and alkyl of 1-6 carbon atoms;
- R.sup.40 and R.sup.50 are independently selected from the group consisting of alkyl of 1-6 carbon atoms;
- R.sup.60 is selected from the group consisting of alkyl of 1-20 carbon atoms, phenyl, alkoxyphenyl, wherein the alkyl group is of 1-10 carbon atoms, and alkylphenyl, wherein the alkyl group is of 1-10 carbon atoms;
- and R.sup.70 is a group of Formula XI
- --(C.dbd.O)R.sup.80 XI
- wherein R.sup.80 is selected from the group consisting of alkyl of 1-20 carbon atoms, phenyl, benzyl, phenyl substituted at any one position with alkyl of 1-10 carbon atoms, chloro, bromo, nitro, or cyano, and benzyl substituted at any one position on the phenyl ring with alkyl of 1-10 carbon atoms, chloro, bromo, nitro or cyano, bathing the support in an aqueous solution with a pH less than about 3, and visualizing the stained protein.
A method of detecting a protein which comprises making a visibly colored complex of the protein with a dye of Formula X ##STR4## wherein R.sup.10, R.sup.20, and R.sup.30 are independently selected from the group consisting of hydrogen and alkyl of 1-6 carbon atoms;
- R.sup.40 and R.sup.50 are independently selected from the group consisting of alkyl of 1-6 carbon atoms;
- R.sup.60 is selected from the group consisting of alkyl of 1-20 carbon atoms, phenyl, alkoxyphenyl, wherein the alkyl group is of 1-10 carbon atoms, and alkylphenyl, wherein the alkyl group is of 1-10 carbon atoms;
- and R.sup.70 is a group of Formula XI
- --(C.dbd.O)R.sup.80 XI
- wherein R.sup.80 is selected from the group consisting of alkyl of 1-20 carbon atoms, phenyl, benzyl, phenyl substituted at any one position with alkyl of 1-10 carbon atoms, chloro, bromo, nitro, or cyano, and benzyl substituted at any one position on the phenyl ring with alkyl of 1-10 carbon atoms, chloro, bromo, nitro or cyano, and detecting the complex by observing the color thereof.
A method of detecting protein in a sample comprising the steps of:
- (a) applying a sample solution comprising a protein to a polyacrylamide or agarose gel;
- (b) separating the protein of the sample solution into protein fractions using electrophoresis;
- (c) staining the protein on the gel with a dye of Formula X ##STR5## wherein R.sup.10, R.sup.20, and R.sup.30 are independently selected from the group consisting of hydrogen and alkyl of 1-6 carbon atoms;
- R.sup.40 and R.sup.50 are independently selected from the group consisting of alkyl of 1-6 carbon atoms;
- R.sup.60 is selected from the group consisting of alkyl of 1-20 carbon atoms, phenyl, alkoxyphenyl, wherein the alkyl group is of 1-10 carbon atoms, and alkylphenyl, wherein the alkyl group is of 1-10 carbon atoms;
- and R.sup.70 is a group of Formula XI
- --(C.dbd.O)R.sup.80 XI
- wherein R.sup.80 is selected from the group consisting of alkyl of 1-20 carbon atoms, phenyl, benzyl, phenyl substituted at any one position with alkyl of 1-10 carbon atoms, chloro, bromo, nitro, or cyano, and benzyl substituted at any one position on the phenyl ring with alkyl of 1-10 carbon atoms, chloro, bromo, nitro or cyano;
- and
- (d) detecting the stained protein by observing the color thereof.
Dependencies are inferred from the claim text.