Labeling proteins with dyes that are insoluble or only sparingly soluble in water
AssigneeBio-Rad Laboratories, Inc.
InventorsMingde Zhu, Lee Olech
Life of this patent
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Abstract
The proteins in a biological sample that is sought to be analyzed for its protein composition by an electrophoretic or chromatographic procedure are coupled to a dye in an unusually efficient manner by combining the sample with a solid dry composition containing the dye, a buffering agent, and in preferred embodiments, a denaturing agent as well. The solid and dry form of the composition prevents the dye from deteriorating or decomposing, and the combination of components in the composition allows the dye to couple to the proteins in a relatively uniform manner with no overstaining of the protein when the composition and the sample are heated together and held at an elevated temperature for a short period of time.
Claims12
A process for the preparation of a protein-containing biological sample for electrophoretic analysis of the protein composition of said sample, said process comprising:
- (a) contacting said sample with a solid dry composition comprising a cyanine succinimidyl ester fluorescent dye whose solubility in water is less than 5 μg/mL at 25° C. and which is capable of coupling to a protein, in the presence of a non-nucleophilic buffering agent and in the absence of organic solvents;
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- (b) while said sample is in contact with said solid dry composition, heating said sample to a non-boiling temperature of at least about 80° C. to cause said fluorescent dye to couple to said protein.
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Dependencies are inferred from the claim text.