Analysis of carbohydrates derivatized with visible dye by high-resolution polyacrylamide gel electrophoresis
AssigneeBoard of Regents, The University of Texas System
InventorLuis S. Haro
Life of this patent
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Abstract
The present invention is related to the field of carbohydrate analysis. More particularly, this invention relates to a simple and inexpensive method for analyzing carbohydrates which can be used to separate mono-, di-, tri- and even poly-saccharides. More specifically, this invention relates to a quantitative and qualitative method for analyzing carbohydrates that may be present in a biological sample by employing a visible dye or chromophore and high resolution polyacrylamide gel electrophoresis.
Claims40
A method of detecting the presence of a carbohydrate in a sample suspected of containing a carbohydrate comprising:
- a) contacting the sample with a non-fluorescent chromophore to produce a labeled carbohydrate, wherein said chromophore comprises 4-amino-1,1′-azobenzene or a sulfonic acid of 4-amino-1,1′-azobenzene;
- b) separating the labeled carbohydrate from any unreacted non-fluorescent chromophore by gel electrophoresis with a gel;
- and
- c) detecting the labeled carbohydrate.
A method of identifying at least two carbohydrates in a sample comprising:
- a) contacting the sample with a non-fluorescent chromophore to produce labeled carbohydrates, wherein said non-fluorescent chromophore is selected from the group consisting of 4-amino-1,1′-azobenzene or a sulfonic acid of 4-amino-1,1′-azobenzene;
- b) separating the labeled carbohydrates by gel electrophoresis with a gel;
- c) halting the gel electrophoresis;
- d) identifying at least one labeled carbohydrate;
- e) resuming gel electrophoresis;
- and
- f) identifying at least one labeled carbohydrate;
- wherein the carbohydrates are identified by comparing migration of the labeled carbohydrates in the gel with the migration of a standard.
A method of identifying an unknown carbohydrate in a sample comprising:
- a) reductively aminating the carbohydrate with 4-amino-1,1′-azobenzene-3,4′-disulfonic acid to produce a non-fluorescent chromophore labeled carbohydrate;
- b) separating the non-fluorescent chromophore labeled carbohydrate from any unreacted 4-amino-1,1′-azobenzene-3,4′-disulfonic acid by polyacrylamide gel electrophoresis with a polyacrylamide gel;
- c) quantifiably detecting the non-fluorescent chromophore labeled carbohydrate with a densitometer;
- and
- d) identifying the carbohydrate by comparing the migration of the non-fluorescent chromophore labeled carbohydrate in the gel with the migration of a standard carbohydrate.
A kit for detecting carbohydrates comprising:
- a non-fluorescent chromophoric labeling reagent comprising 4-amino-1,1′-azobenzene or a sulfonic acid of 4-amino-1,1′-azobenzene;
- a precast polyacrylamide gel;
- an electrophoretic buffer;
- and
- a carbohydrate standard.
Dependencies are inferred from the claim text.