title: Simplified methods for isolation of ubiquitin from erythrocytes. Generation of ubiquitin polymers.
abstract: 1. Ubiquitin has been isolated from bovine erythrocytes by procedures in which the hemoglobin was removed by denaturation with either ethanol-chloroform mixtures or by heating. 2. The proteins soluble to the denaturation step were removed by 3% sodium trichloroacetate (TCA) at pH 2.0-2.5 or by 5% TCA. 3. Ubiquitin was isolated in relatively high yield from the TCA insoluble fraction by use of single ion-exchange chromatographic and gel permeation steps. 4. Ubiquitin shows relatively little cross-linking upon treatment with glutaraldehyde or with dimethyl suberimidate. Heating of the glutaraldehyde treated material in 4 M guanidine, however, leads to marked aggregation. 5. The polymers of ubiquitin react strongly with antibody in an immunoblot assay.
authors.affiliation: Max-Planck-Institut für Zellbiologie, Ladenburg, West Germany.
authors.forename: H F
authors.initials: HF
authors.lastname: Deutsch
mesh_terms: D000818:Animals; D002417:Cattle; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004912:Erythrocytes / Q000378:metabolism*; D012967:Sodium Dodecyl Sulfate; D013552:Swine; D014452:Ubiquitins / Q000097:blood*
chemical_list: D014452:Ubiquitins; D012967:Sodium Dodecyl Sulfate