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US 5,705,649 AGranted
Protein staining compositions and methods
AssigneePromega Corporation
InventorsJohn W. Shultz, David L. Leland
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AssigneePromega Corporation
InventorsJohn W. Shultz, David L. Leland
Estimating the term…
The present invention provides novel dyes for staining proteins, particularly those in electrophoretic gels, and detecting the proteins with high sensitivity. The dyes of the invention are derivatives of known dyes that are more hydrophobic than the corresponding, known dyes and, in electrophoretic gels, including polyacrylamide gels with sodium dodecyl sulfate, form easily detectable, non-covalent complexes with proteins. In some cases, the corresponding, known dyes bind to protein covalently. The complexes of protein with some dyes of the invention can be detected with high sensitivity simply by observation of the color of the complexes by eye. The complexes of protein with other dyes of the invention are detected by observation of the fluorescence of the complexes. Dyes of the invention include derivatives of Coomassie.RTM. dyes, acid blue 25, dansyl chloride, fluorescein, 2-methyl-2,4-diphenyl-3(2H)-furanone, and (7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino alkanoates. The invention also provides quantitative and qualitative methods for detecting proteins which comprise staining the proteins with a dye of the invention.
A dye of Formula of XCI ##STR75## wherein R.sup.82 is --(CH.sub.2).sub.n --, n having the value of 1-20, bb is 1, R.sup.83 is hydrogen or alkyl of 1-20 carbons, phenyl, or benzyl, and R.sup.84 is, independently of R.sup.83, hydrogen, alkyl of 1-20 carbons, phenyl or benzyl, provided that at least one of R.sup.83 and R.sup.84 is other than hydrogen, or a salt thereof.
A method of detecting a protein which comprises (A) making a complex of the protein with a dye of Formula XCI ##STR76## wherein R.sup.82 is --(CH.sub.2).sub.n --, n having the value of 1-20, bb is 0 or 1, R.sup.83 is hydrogen or alkyl of 1-20 carbons, phenyl, or benzyl, and R.sup.84 is, independently of R.sup.83, hydrogen, alkyl of 1-20 carbons, phenyl or benzyl, provided that at least one of R.sup.83 and R.sup.84 is other than hydrogen, and (B) detecting the complex by observing the color or the fluorescence of the complex due to the presence of the dye of Formula XCI in the complex.
A method of detecting protein in a sample comprising the steps of:
Dependencies are inferred from the claim text.