Two-dimensional electrophoretic separation of carbohydrates

Chemistry: electrical and wave energy – Processes and products – Electrostatic field or electrical discharge

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2041828, 204299R, 536127, G01N 2726

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active

050947400

ABSTRACT:
Separation methodology is disclosed which allows for the separation of mixtures of carbohydrates into highly resolved detectable bands of carbohydrates. The method involves first reacting a mixture of carbohydrates with charge generating moieties which are capable of fluorescing such as 1-amino-4-naphthalene sulfonic acid (ANSA) to form carbohydrate conjugates. The conjugates are subjected to a first-dimensional gel electrophoresis in a first direction to provide separate bands of carbohydrates in the gel. A band in the gel is removed and subjected to second-dimensional electrophoresis in a second direction which is substantially perpendicular to the first direction. More specific bands of more highly resolved carbohydrates are then formed in the second-dimensional gel. The more specific bands within the second-dimensional gel are then electro-blotted onto a substrate surface and can be viewed in extremely small amounts due to the fluorescent capability of the ANSA when viewed under ultraviolet light.

REFERENCES:
patent: 4109075 (1978-08-01), Deaton
patent: 4305799 (1981-12-01), Schwartz et al.
patent: 4666581 (1987-05-01), Itoh et al.
patent: 4975165 (1990-12-01), Brandley et al.

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