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Title: Electrophoretic extraction of proteins from two-dimensional electrophoresis gel spots

Abstract

After two-dimensional electrophoresis of proteins or the like, resulting in a polyacrylamide gel slab having a pattern of protein gel spots thereon, an individual protein gel spot is cored out from the slab, to form a gel spot core which is placed in an extraction tube, with a dialysis membrane across the lower end of the tube. Replicate gel spots can be cored out from replicate gel slabs and placed in the extraction tube. Molten agarose gel is poured into the extraction tube where the agarose gel hardens to form an immobilizing gel, covering the gel spot cores. The upper end portion of the extraction tube is filled with a volume of buffer solution, and the upper end is closed by another dialysis membrane. Upper and lower bodies of a buffer solution are brought into contact with the upper and lower membranes and are provided with electrodes connected to the positive and negative terminals of a dc power supply, thereby producing an electrical current which flows through the upper membrane, the volume of buffer solution, the agarose, the gel spot cores and the lower membrane. The current causes the proteins to be extracted electrophoretically from the gel spot cores, somore » that the extracted proteins accumulate and are contained in the space between the agarose gel and the upper membrane. 8 figs.« less

Inventors:
; ;
Issue Date:
Research Org.:
Argonne National Lab., IL (USA)
OSTI Identifier:
6736991
Patent Number(s):
-US-A6092924
Assignee:
ANL; ERA-14-001854; EDB-88-186804
DOE Contract Number:  
W-31109-ENG-38
Resource Type:
Patent
Resource Relation:
Other Information: Portions of this document are illegible in microfiche products
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; PROTEINS; EXTRACTION; TWO-DIMENSIONAL ELECTROPHORESIS; OPTIMIZATION; GELS; INVENTIONS; SOLIDIFICATION; COLLOIDS; DISPERSIONS; ELECTROPHORESIS; ORGANIC COMPOUNDS; PHASE TRANSFORMATIONS; SEPARATION PROCESSES 550200* -- Biochemistry

Citation Formats

Zhang, Jian-Shi, Giometti, C.S., and Tollaksen, S.L. Electrophoretic extraction of proteins from two-dimensional electrophoresis gel spots. United States: N. p., 1987. Web.
Zhang, Jian-Shi, Giometti, C.S., & Tollaksen, S.L. Electrophoretic extraction of proteins from two-dimensional electrophoresis gel spots. United States.
Zhang, Jian-Shi, Giometti, C.S., and Tollaksen, S.L. Fri . "Electrophoretic extraction of proteins from two-dimensional electrophoresis gel spots". United States.
@article{osti_6736991,
title = {Electrophoretic extraction of proteins from two-dimensional electrophoresis gel spots},
author = {Zhang, Jian-Shi and Giometti, C.S. and Tollaksen, S.L.},
abstractNote = {After two-dimensional electrophoresis of proteins or the like, resulting in a polyacrylamide gel slab having a pattern of protein gel spots thereon, an individual protein gel spot is cored out from the slab, to form a gel spot core which is placed in an extraction tube, with a dialysis membrane across the lower end of the tube. Replicate gel spots can be cored out from replicate gel slabs and placed in the extraction tube. Molten agarose gel is poured into the extraction tube where the agarose gel hardens to form an immobilizing gel, covering the gel spot cores. The upper end portion of the extraction tube is filled with a volume of buffer solution, and the upper end is closed by another dialysis membrane. Upper and lower bodies of a buffer solution are brought into contact with the upper and lower membranes and are provided with electrodes connected to the positive and negative terminals of a dc power supply, thereby producing an electrical current which flows through the upper membrane, the volume of buffer solution, the agarose, the gel spot cores and the lower membrane. The current causes the proteins to be extracted electrophoretically from the gel spot cores, so that the extracted proteins accumulate and are contained in the space between the agarose gel and the upper membrane. 8 figs.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {1987},
month = {9}
}