Tiselius electrophoresis method

Revs 24, 271(1939) (Electrophoresis of proteins by Tiselius Method) 2a) Ibid, JACS 62, 703(1940) (Interpretation of simple electrophoretic patterns) 3) H.A. Abramson,... 

The unidimensional type of paper electrophoresis is an extension of free boundary electrophoresis, the method developed by Tiselius (Tl). There are several differences between the two systems. One is the presence of a substrate (supporting medium) as anticonvectional medium during the electrophoretic separation. Another important difference is the starting point. In paper electrophoresis the entire load of material due to be separated is collected on the starting line, whereas in free boundary electrophoresis the material is present in equal concentration over one leg of the electrophoretic cell. Fortunately these differences simplify the qualitative and quantitative appraisal of separation after the run on paper, and for practical work both prove to be true inherent qualities and go far to account for the success of the method (Kl, VI, Wl). 

Chromatography of polysaccharides on silica gel and on carbon columns has been used to a limited extent. The use of ion-exchange resins has been investigated by Deuel and coworkers. The colunms had low capacities, and part of the material appeared to be irreversibly adsorbed. Deuel and coworkers, however, have reported better results when columns of cationic derivatives of cellulose, for instance (diethylaminoethyl)-cellulose, are used. Electrophoresis both by the Tiselius method on columns and on filter paper or glass-fiber sheets can give good separa-... 

A number of years later. Smithies Introduced starch gels in the 1950 s (, ). His technique of zone electrophoresis with starch gel as the supporting medium gave a resolving power equal, if not greater, to that of the Tiselius method. Shortly thereafter,... 

The Tiselius method of determining electrophoresis has been especially important in the investigation of proteins and other hydrophilic colloids One of its essential features is the possibility of determining electrophoresis in a mixture of different proteins The Schlicren picture then contains two or more different humps Another interesting feature of the method is the possibility of using it for the separations of mixtures of proteins. 

The origins, principles, methods, and modes of capillary electrophoresis (CE) are discussed. Massive application of electrophoresis methods started after Tiselius s moving boundary method that was optimized by the use of paper or a gel as a semiconducting medium. The applications of paper and gel electrophoresis were situated mostly in the biochemical environment for the analysis of proteins, amino... 

With the study of the migration of hydrogenium ions (H ) in a phenolphthalein gel by Lodge in 1886 and the description of the migration of ions in saline solutions by Kohlraush in 1897, a basis was set for the development of a new separation technique that we know today as electrophoresis. Indeed, several authors applied the concepts introduced by Lodge and Kohlraush in their methods and when Arne Tiselius reported the separation of different serum proteins in 1937, the approach called electrophoresis was recognized as a potential analytical technique. Tiselius received the Nobel Prize in Chemistry for the introduction of the method called moving boundary electrophoresis. ... 

A Tiselius. Moving boundary method of studying the electrophoresis of proteins. Nova Acta Reg Soc Uppsal Ser IV 7 1-107, 1930. 

A. Tiselius, The moving boundary method of studying the electrophoresis of proteins, Ph.D. Thesis, Nova acta regiae societatis scientiarum, Ser. IV, Vol. 17, No. 4, Uppsala, Sweden Almqvist Wiksell (1930), 1-107. 

Arne Tiselius announced his electrophoresis apparatus, which was enormous both in size and expense, in 1937. Electrophoresis was designed for the separation and investigation of proteins, and the technique has had its greatest impact in biology and biochemistry.174 The development of electrophoretic methods after Tiselius has been... 

Aug. 10,1902, Stockholm, Sweden - Oct. 29,1971, Uppsala, Sweden) Tiselius studied chemistry in Uppsala and joined the laboratory of -> Svedberg in 1925. In 1930 he received the Ph.D. for his thesis entitled The moving-boundary method of studying the electrophoresis of proteins . In 1937, a research professorship was established for Tiselius. His fundamental work in developing -> electrophoresis was encouraged and supported by Svedberg. In 1948 he was awarded the Nobel Prize in Chemistry for his achievements in electrophoresis and adsorption analysis. 

Tiselius, Arne W. K. (1902-1971). A Swedish biochemist who won the Nobel Prize for chemistry in 1948. Renowned for research in separation methods of biochemical matter, in particular electrophoresis and chromatography. Work also involved virus isolation and synthesis of blood plasma. He earned degrees from the University of Upsala and Princeton University, as well as a multitude of honorary degrees. 

Electrophoresis is a separation method based on the differential rates of migration of charged species in an applied dc electric field. This separation technique for macrosize samples was first developed by the Swedish chemist Arne Tiselius in the 1930s for the study of semm proteins he was awarded the 1948 Nobel Prize for his work. 

The moving boundary method for investigating electrophoresis has, however, been improved by Tiselius to the extent that it has become a powerful tool of investigation, particularly of proteins. Tiselius 10 early work is outlined in his dissertation which includes a valuable discussion of the conditions governing the movement of a boundary. A particularly important experimental detail is that the composition of the solution in which the colloid is suspended should be, as nearly as possible, of the same composition as the solution with which it is in contact at the boundary, otherwise boundary anomalies will be present. These will cause the particles of colloid to move abnormally fast or slow, and may even give rise to additional boundaries. 

Arne Tiselius, "The Moving Boundary Method of Studying the Electrophoresis of PrO-... 

Development of electrophoretic protein separation techniques have been paralleled by improvements in protein detection methods. Protein detection in early electrophoretic applications, utilizing electrophoretic separations of solutions or colloidal suspensions from about 1816 to 1937, was limited to direct visualization of proteins coated onto microspheres, or studies of naturally colored proteins such as hemoglobin, myoglobin, or ferritin <1-4). An increase in sensitivity and the ability to detect non-colored proteins was achieved by the use of the specific absorption, by proteins, of ultraviolet light. This detection technique permitted Tiselius,in 1937, to demonstrate the quantitative electrophoretic separation of ovalbumin, serum globulin fractions and Bence Jones proteins (S). Tiselius also employed the shadows, or schlieren, created by the boundaries, due to the different concentrations of proteins in the electrophoretic system to detect protein position and concentration ( ). These detection methods served as the main methods for protein detection in the liquid electrophoresis systems. However,... 

Another purified enzyme preparation which produces laminaripentaose from insoluble laminarin and from heat-treated pachyman is produced by a strain of Arthrobacter luteus (100,101,102) when grown on yeast cells or / -(1 —>3)-glucan. The enzyme, which was named Zymolase (also referred to as Zymolyase) appeared to be homogeneous by electrophoresis in a Tiselius apparatus and by ultracentrifugation. The molecular weight of the enzyme was estimated from ultracentrifugation to be ca. 20,500. The optimum pH for lysis of viable yeast cells was 7.5. The optimum temperature was 35°C. The optimum pH for heat-treated pachyman hydrolysis was 6.5, and the optimum temperature was 45°C. A Lineweaver-Burk plot with heat-treated pachyman yielded a Km value of 0.04% when the solubilized carbohydrate was assayed by the phenol-sulfuric acid method. Zymolase lost all its activity after incubation at 60°C for 5 min. 

This technique, although described in the literature, has been given little attention. Isoelectric focusing without stabilizing media can be done in an apparatus similar to Tiselius free zone electrophoresis [152]. Separation itself occurs in a horizontal quartz tube that is rotated at 40 rpm to counteract convective forces [153]. The pH gradient is evolved without the addition of ampholytes, however this method leads to either too steep or too shallow gradients and is therefore not practically applicable. Another alternative method for free solution isoelectric focusing is the application of polyethylene coils however this procedure makes use of ampholines [154]. 

The first important experiments on electrophoresis were carried out by the Swedish physical chemist Arne Wilhelm Kaurin Tiselius (1902-1971), who in 1937 made electrophoresis a powerful technique for studying mixtures of proteins. He devised a special type of U-tube, shown schematically in Figure 11.20, along which the protein molecules move under the influence of an electric potential. Different proteins will move at different speeds. The tube consisted of portions fitted together at ground-glass joints, so that one of a mixture of proteins could be isolated in one chamber. Optical methods are used to determine the quantity of each protein present in the mixture. 

---