Electrophoretic extraction of proteins from two-dimensional electrophoresis gel spots

Zhang, Jian-Shi; Giometti, C S; Tollaksen, S L

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

Full Record
Other Related Research

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 »« less

Inventors:: Zhang, Jian-Shi; Giometti, C S; Tollaksen, S L

Issue Date:: 1987-09-04

Research Org.:: Argonne National Lab. (ANL), Argonne, IL (United States)

OSTI Identifier:: 6736991

Patent Number(s):: 6092924

Assignee:: ANL; ERA-14-001854; EDB-88-186804

Patent Classifications (CPCs):: G - PHYSICS G01 - MEASURING G01N - INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES

Show more

G - PHYSICS G01 - MEASURING G01N - INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
G01N22/04 - Investigating moisture content

Show less

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.

Copy to clipboard


                    Zhang, Jian-Shi, Giometti, C S, & Tollaksen, S L. Electrophoretic extraction of proteins from two-dimensional electrophoresis gel spots.  United States.

Copy to clipboard


                    Zhang, Jian-Shi, Giometti, C S, and Tollaksen, S L. Fri .  
        "Electrophoretic extraction of proteins from two-dimensional electrophoresis gel spots".  United States.

Copy to clipboard


                    
@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}

}

Copy to clipboard

Patent:

Search for the full text at the U.S. Patent and Trademark Office

Save / Share:

Export Metadata

Save to My Library

Similar Records in DOE Patents and OSTI.GOV collections:

Electrophoretic extraction of proteins from two-dimensional electrophoresis gel spots

Patent Zhang, Jian-Shi [Shanghai, CN]; Giometti, Carol S [Glenview, IL]; Tollaksen, Sandra L [Montgomery, IL]

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 uppermore »« less
Full Text Available
Two-dimensional peptide mapping of protease-digested proteins from two-dimensional electrophoresis gel spots

Conference Zhang, J S; Giometti, C S; Tollaksen, S L

A simple procedure is described to extract proteins from polyacrylamide gels and subsequently separate them after partial proteolysis. This method is useful for the biochemical characterization of individual proteins in two-dimensional gel patterns and for the identification of both similarities and differences between a pair of related protein spots. Because the number of gel spots to be extracted and the extraction volume can be adjusted in this procedure, differences in the primary structure of both abundant and minor proteins can be investigated. By extracting protein from 10 or more gel spots, enough protein should be obtained to allow amino acidmore »« less
Cytoskeletal proteins from human skin fibroblasts, peripheral blood leukocytes, and a lymphoblastoid cell line compared by two-dimensional gel electrophoresis

Journal Article Giometti, C S; Willard, K E; Anderson, N L - Clin. Chem. (Winston-Salem, N.C.); (United States)

Differences in proteins between cells grown as suspension cultures and those grown as attached cultures were studied by comparing the proteins of detergent-resistant cytoskeletons prepared from peripheral blood leukocytes and a lymphoblastoid cell line (GM607) (both grown as suspension cultures) and those of human skin fibroblasts (grown as attached cultures) by two-dimensional gel electrophoresis. The major cytoskeletal proteins of the leukocytes were also present in the protein pattern of GM607 cytoskeletons. In contrast, the fibroblast cytoskeletal protein pattern contained four groups of proteins that differed from the patterns of the leukocytes and GM607. In addition, surface labeling of GM607 andmore »« less
Development in electrophoresis: instrumentation for two-dimensional gel electrophoresis of protein separation and application of capillary electrophoresis in micro-bioanalysis

Thesis/Dissertation Xu, Aoshuang [Iowa State Univ., Ames, IA (United States)]

This dissertation begins with a general introduction of topics related to this work. The following chapters contain three scientific manuscripts, each presented in a separate chapter with accompanying tables, figures, and literature citations. The final chapter summarizes the work and provides some prospective on this work. This introduction starts with a brief treatment of the basic principles of electrophoresis separation, followed by a discussion of gel electrophoresis and particularly polyacrylamide gel electrophoresis for protein separation, a summary of common capillary electrophoresis separation modes, and a brief treatment of micro-bioanalysis application of capillary electrophoresis, and ends with an overview of proteinmore » conformation and dynamics.« less
DOI: https://doi.org/10.2172/1342558

Full Text Available
Comparison of protein patterns of xrs-5, a radiosensitive Chinese hamster ovary cell line, and CHO-K1, its radioresistant parent, using two-dimensional gel-electrophoresis

Conference Kramer, J M [Miami Univ., Oxford, OH (USA). Dept. of Zoology]

X-ray sensitive strains of Chinese hamster ovary cell lines have been used to analyze radiation repair mechanisms. One cell line, xrs-5, has been shown to be very sensitive to ionizing radiation and radical forming chemical mutagens. This sensitivity is thought to be a result a mutation in the DNA double strand break (DSB) repair mechanism, and its characterization has been a goal of several repair mechanism studies. Using two-dimensional gel electrophoresis, we have detected a protein (MW approximately 55KD) in the DNA/Nuclear Matrix (nucleoid) cell fraction of CHO-Kl cells that is absent in the nucleoid fraction of xrs-5. This proteinmore »« less
Full Text Available

Similar Records