Difference detection methods using matched multiple dyes
First Claim
1. A set of matched luminescent dyes comprising at least two different dyes which in use covalently bind to proteins within an extract of proteins from at least two cell samples, wherein the dyes within said set:
- (a) have a matched net charge which will maintain the overall net charge of the proteins upon such covalent binding and matched ionic and pH characteristics whereby relative migration of a protein labeled with any one of said dyes is the same as relative migration of said protein labeled with another dye in said set; and
(b) each emit luminescent light at a wavelength that is sufficiently different from the emitted luminescent light of remaining dyes in said set to provide a detectably different light signal from the light signal of said remaining dyes in said set.
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Abstract
A process and a kit are provided for detecting differences in two or more samples of protein, including proteins bearing post-translational modifications and peptides. Proteins are prepared, for example, from each of a different group of cell samples or body fluid samples to be compared. Each protein extract is labeled with a different one of a luminescent dye from a matched set of dyes. The matched dyes have generally the same ionic and pH characteristics but emit light at different wavelengths to exhibit a different color upon luminescence detection. The labeled protein extracts are mixed together and separated together by electrophoresis or a chromatographic method. The separation is observed to detect proteins unique to one sample or present in a greater ratio in one sample than in the other. Those unique or excess proteins will fluoresce the color of one of the dyes used. Proteins common to each sample migrate together and fluoresce the same.
28 Citations
44 Claims
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1. A set of matched luminescent dyes comprising at least two different dyes which in use covalently bind to proteins within an extract of proteins from at least two cell samples, wherein the dyes within said set:
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(a) have a matched net charge which will maintain the overall net charge of the proteins upon such covalent binding and matched ionic and pH characteristics whereby relative migration of a protein labeled with any one of said dyes is the same as relative migration of said protein labeled with another dye in said set; and (b) each emit luminescent light at a wavelength that is sufficiently different from the emitted luminescent light of remaining dyes in said set to provide a detectably different light signal from the light signal of said remaining dyes in said set. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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15. A set of matched luminescent dyes comprising at least two different dyes which in use covalently bind to proteins within a mixture of proteins from each of at least two samples, wherein the dyes within said set:
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(a) have matched ionic and pH characteristics whereby relative migration of a protein labeled with any one of said dyes is the same as relative migration of said protein labeled with another dye in said set; and (b) each emit luminescent light at a wavelength that is sufficiently different from the emitted luminescent light of remaining dyes in said set to provide a detectably different light signal from the light signal of said remaining dyes in said set. - View Dependent Claims (16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44)
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Specification