Two-dimensional electrophoresis: agarose gel isotachophoresis followed by sodium dodecyl sulphate—polyacrylamide electrophoresis

1991 ◽  
Vol 545 (2) ◽  
pp. 343-347 ◽  
Author(s):  
Fernando Acevedo ◽  
Zere Goitom
Keyword(s):  
Sodium Dodecyl Sulphate ◽  
Sodium Dodecyl ◽  
Agarose Gel ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Polyacrylamide Electrophoresis ◽  
Dimensional Electrophoresis

scholarly journals A novel system for the two-dimensional electrophoresis of membrane proteins

1977 ◽  
Vol 163 (1) ◽  
pp. 165-168 ◽  
Author(s):  
A G Booth
Keyword(s):  
Membrane Proteins ◽  
Sodium Dodecyl Sulphate ◽  
Chloral Hydrate ◽  
Sodium Dodecyl ◽  
Two Dimensions ◽  
Two Dimensional ◽  
Organic Base ◽  
Two Dimensional Electrophoresis ◽  
Novel Technique ◽  
Dimensional Electrophoresis

Membrane proteins were resolved in two dimensions by a novel technique that uses discontinuous electrophoresis in both directions. After electrophoresis in the first direction in chloral hydrate, the membrane proteins were further resolved by a novel system that used organic-base dodecyl sulphates to stack and then resolve them. This latter system has several advantages over conventional electrophoresis in sodium dodecyl sulphate, notably that it avoids the production of artifacts generated by other systems.

scholarly journals Red cell proteins. I. Two-dimensional mapping of human erythrocyte lysate proteins

Blood ◽  
1979 ◽  
Vol 53 (6) ◽  
pp. 1121-1132 ◽  
Author(s):  
JJ Edwards ◽  
NG Anderson ◽  
SL Nance ◽  
NL Anderson
Keyword(s):  
Human Erythrocyte ◽  
Sodium Dodecyl ◽  
Deae Cellulose ◽  
Cell Protein ◽  
Red Cell ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Mapping Techniques ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Dimensional Electrophoresis

Abstract Human erythrocyte lysate proteins were resolved into over 250 discrete spots by two-dimensional electrophoresis using isoelectric focusing in the first dimension and electrophoresis in the presence of sodium dodecyl sulfate, (SDS) in the second. The overwhelming excess of hemoglobin has made such analyses difficult in the past. However, with the ISO-DALT two-dimensional electrophoresis system, large numbers of red cell proteins can be mapped in the presence of hemoglobin. When hemoglobin and several other major proteins are removed by adsorption to DEAE-cellulose, additional minor components are seen, giving a total of over 275. With the use of purified preparations, the map positions of five cell enzymes or their subunits were determined: pyruvate kinase, catalase, glucose-6-phosphate dehydrogenase, hypoxanthine phosphoribosyltransferase, and carbonic anhydrase. The mapping techniques described complement and extend those traditionally used to find human red cell protein variants.

Evaluation of lipoproteins and apolipoproteins in serum of a Tangier patient by micro-scale two-dimensional electrophoresis.

1987 ◽  
Vol 33 (4) ◽  
pp. 468-472 ◽  
Author(s):  
T Manabe ◽  
S Visvikis ◽  
M F Dumon ◽  
M Clerc ◽  
G Siest
Keyword(s):  
Molecular Mass ◽  
Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Disease Patient ◽  
High Density Lipoproteins ◽  
Two Dimensional ◽  
High Concentration ◽  
Two Dimensional Electrophoresis ◽  
Micro Scale ◽  
Dimensional Electrophoresis

Abstract We examined lipoproteins and apolipoproteins in serum of a Tangier-disease patient. We used three different techniques of micro-scale two-dimensional electrophoresis: (a) no denaturants; (b) with sodium dodecyl sulfate (SDS) used only in the slab gel electrophoresis; (c) and with urea and a detergent used in isoelectric focusing and with SDS in slab gel electrophoresis. By technique a, an extremely low concentration of high-density lipoproteins (HDL) in the Tangier serum was seen, and lipoproteins that cannot form HDL complexes were detected as multiple spots in the acidic (pl 4 approximately 5) and relatively low apparent molecular mass (20,000 approximately 80,000) region. By technique b, Tangier low-molecular-mass lipoproteins were dissociated into their constituent apolipoproteins, and we observed a higher proportion of apoC-III, together with lower proportions of apoA-I and apoA-II, than in the normal HDL fraction. Technique c showed the total content of apolipoproteins in the whole Tangier serum, as several workers have reported. The presence of low-molecular-mass lipoproteins and a high concentration of apoC-III in this lipoprotein fraction characterized the Tangier serum.

THE NATURE OF TWO COMPONENTS OF PIG MUCOSAL HEPARIN, SEPARATED BY ELECTROPHORESIS IN AGAROSE GEL

1967 ◽  
Vol 45 (7) ◽  
pp. 1015-1020 ◽  
Author(s):  
Alan A. Horner
Keyword(s):  
Gel Electrophoresis ◽  
Nitrous Acid ◽  
Anticoagulant Activity ◽  
Thin Layers ◽  
Agarose Gel ◽  
Two Dimensional ◽  
Agarose Gel Electrophoresis ◽  
Uronic Acids ◽  
Two Dimensional Electrophoresis ◽  
Dimensional Electrophoresis

Two components of pig mucosal heparin were separated by agarose-gel electrophoresis and recovered from the gels. The slower moving component had the higher anticoagulant activity and intrinsic viscosity. The faster moving component had the higher sulfate : carboxyl ratio. Each component was degraded by self-hydrolysis to N-desulfated heparin, deaminated with nitrous acid, and hydrolyzed with formic acid. Two-dimensional electrophoresis and chromatography on thin layers of cellulose showed that each component had two uronic acids with the characteristics of glucuronic and iduronic acids.

scholarly journals Red cell proteins. I. Two-dimensional mapping of human erythrocyte lysate proteins

Blood ◽  
1979 ◽  
Vol 53 (6) ◽  
pp. 1121-1132
Author(s):  
JJ Edwards ◽  
NG Anderson ◽  
SL Nance ◽  
NL Anderson
Keyword(s):  
Human Erythrocyte ◽  
Sodium Dodecyl ◽  
Deae Cellulose ◽  
Cell Protein ◽  
Red Cell ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Mapping Techniques ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Dimensional Electrophoresis

Human erythrocyte lysate proteins were resolved into over 250 discrete spots by two-dimensional electrophoresis using isoelectric focusing in the first dimension and electrophoresis in the presence of sodium dodecyl sulfate, (SDS) in the second. The overwhelming excess of hemoglobin has made such analyses difficult in the past. However, with the ISO-DALT two-dimensional electrophoresis system, large numbers of red cell proteins can be mapped in the presence of hemoglobin. When hemoglobin and several other major proteins are removed by adsorption to DEAE-cellulose, additional minor components are seen, giving a total of over 275. With the use of purified preparations, the map positions of five cell enzymes or their subunits were determined: pyruvate kinase, catalase, glucose-6-phosphate dehydrogenase, hypoxanthine phosphoribosyltransferase, and carbonic anhydrase. The mapping techniques described complement and extend those traditionally used to find human red cell protein variants.

An attempt to resolve all the various proteins in a single human cell type by two-dimensional electrophoresis: I. Extraction of all cell proteins.

1984 ◽  
Vol 30 (12) ◽  
pp. 2014-2020 ◽  
Author(s):  
J Klose ◽  
E Zeindl
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Human Cell ◽  
Sodium Dodecyl ◽  
Two Dimensional ◽  
Cell Type ◽  
Two Dimensional Electrophoresis ◽  
Extraction Procedures ◽  
Dodecyl Sulfate ◽  
Protein Classes ◽  
Dimensional Electrophoresis

Abstract A concept is presented for estimation of the total number of different proteins in a single human cell type (exemplified here by Hep cells) by use of two-dimensional electrophoresis (2DE). This concept includes three problems, the first, investigated in this study, being the transfer of all protein species of the cells into a sample useful for separation by 2DE. Five different extraction media containing--in various combinations--urea, Nonidet P-40, Zwittergent, mercaptoethanol, dithiothreitol, and sodium dodecyl sulfate were used step by step in three different extraction procedures to extract the cell proteins. The amount of radiolabeled proteins in each extract was measured. Each extract was subjected to 2DE. From the total mass of cell proteins, 99.99% could be extracted in two steps: 96% were extracted with urea/beta-mercaptoethanol solution, the remaining 4% with sodium dodecyl sulfate/urea/beta-mercaptoethanol solution. A special class of proteins assumed to be present in the latter fraction was not detected. Thus this fraction can be omitted from the further analysis of all cell proteins by 2DE. Protein classes that possibly remain undetected by the described extraction procedures are mentioned.

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