title: Preparative high-yield electroelution of proteins after separation by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and its application to analysis of amino acid sequences and to raise antibodies.
abstract: A method for the preparative high-yield electroelution of proteins from sodium dodecyl sulphate (SDS) polyacrylamide gel strips was established. The method consisted of SDS-polyacrylamide gel electrophoresis, detection of proteins with sodium acetate and electrophoretic elution at 200 V for 3 h by utilizing a horizontal flat-bed gel electrophoresis apparatus. Standard proteins with molecular masses of 14-66 kilodalton (cytochrome c, aldolase, ovalbumin and bovine serum albumin) were recovered with an average yield of 73.6 +/- 2.3%. A membrane-bound protein, rat skeletal muscle Ca(2+)-ATPase (100 kilodalton) was also well recovered (over 60%). This method was applicable to the purification of proteins required for N-terminal amino acid sequencing and to raise antibodies.
authors.affiliation: Faculty of Pharmaceutical Sciences, Okayama University, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Ohhashi
authors.forename: C
authors.initials: C
authors.lastname: Moritani
authors.forename: H
authors.initials: H
authors.lastname: Andoh
authors.forename: S
authors.initials: S
authors.lastname: Satoh
authors.forename: S
authors.initials: S
authors.lastname: Ohmori
authors.forename: F
authors.initials: F
authors.lastname: Lottspeich
authors.forename: M
authors.initials: M
authors.lastname: Ikeda
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000252:Calcium-Transporting ATPases / Q000032:analysis / Q000276:immunology; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003574:Cytochrome c Group / Q000032:analysis / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005634:Fructose-Bisphosphate Aldolase / Q000032:analysis / Q000276:immunology; D015151:Immunoblotting; D008969:Molecular Sequence Data; D010047:Ovalbumin / Q000032:analysis / Q000276:immunology; D011506:Proteins / Q000032:analysis* / Q000276:immunology*; D011817:Rabbits; D051381:Rats; D012710:Serum Albumin, Bovine / Q000032:analysis / Q000276:immunology; D012967:Sodium Dodecyl Sulfate
chemical_list: D000906:Antibodies; D003574:Cytochrome c Group; D011506:Proteins; D012710:Serum Albumin, Bovine; D012967:Sodium Dodecyl Sulfate; D010047:Ovalbumin; D005634:Fructose-Bisphosphate Aldolase; D000252:Calcium-Transporting ATPases