A VERSATILE HIGH-VOLTAGE PAPER ELECTROPHORESIS APPARATUS AND ITS APPLICATION TO COMPLEX BIOLOGICAL MATERIALS

1961 ◽  
Vol 39 (8) ◽  
pp. 1313-1324 ◽  
Author(s):  
Arthur E. Pasieka
Keyword(s):  
Amino Acids ◽  
High Voltage ◽  
Filter Paper ◽  
Heat Exchangers ◽  
Cooling System ◽  
Paper Electrophoresis ◽  
Biological Materials ◽  
Heat Conductance ◽  
Cooling Plate ◽  
External Cooling

Improvements on a high-voltage paper electrophoresis apparatus are described. The major improvements consist of a simplified cooling system and modification of the cooling plate to accommodate filter paper sheets 24 × 48 in. in size. Two methods, both employing solid heat exchangers, have been devised for the dissipation of Joule's heat: (a) external cooling, employing a Freon 12 circulating coolant and (b) no external cooling, but with heat conductance and radiation by means of an aluminum plate and metal rail.The application of this modified apparatus to the separation of amino acids and peptides from complex biological materials is illustrated.

A TECHNIQUE OF PREPARATIVE PAPER CHROMATOGRAPHY

1966 ◽  
Vol 44 (2) ◽  
pp. 149-153 ◽  
Author(s):  
A. E. Pasieka ◽  
J. E. Logan
Keyword(s):  
Amino Acids ◽  
Paper Chromatography ◽  
Filter Paper ◽  
Biological Materials ◽  
High Salt ◽  
Salt Medium ◽  
Preparative Scale ◽  
High Concentrations ◽  
Total Amino Acids

The use of a solvent redevelopment technique enables the separation of amino acids from complex biological materials in the presence of high concentrations of salts. By conventional chromatography, 1.17 mg of total amino acids have been separated from the high salt medium M 150. The preparative technique as described here has separated amounts as great as 1.17 g and the patterns are essentially the same as for the analytical types. The separations are effected by four or more successive 15- to 20-hour solvent developments with drying between each solvent stage before the staining of chromatograms or isolation of particular bands. The results of these solvent developments on the preparative scale are illustrated with photographs of actual chromatograms. This technique requires thick filter paper sheets up to 4 ft in length for analytical, and particularly for preparative, chromatograms.

INCORPORATION OF C14INTO BACTERIAL PEPTIDES

1958 ◽  
Vol 4 (6) ◽  
pp. 633-648 ◽  
Author(s):  
G. E. Connell ◽  
R. W. Watson
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Acid Hydrolysis ◽  
High Voltage ◽  
Basic Amino Acid ◽  
Paper Electrophoresis ◽  
Total Radioactivity ◽  
Before And After ◽  
Basic Peptides ◽  
Peptide Fractions

Alcoholic extracts of log-phase cells of Pseudomonas hydrophila grown on glucose as the sole carbon source were concentrated, extracted with ether, and dialyzed. After fractionation of the permeates by high voltage paper electrophoresis, five to seven ninhydrin-positive acidic, one neutral, and several basic bands appeared. As many as seven ninhydrin-positive basic bands have been found in some preparations, although they have been absent in others. Photometric analyses on eluates before and after acid hydrolysis indicated the presence of acidic, neutral, and basic peptides. Amino acid compositions of 28 of these peptides, separated by a combination of high voltage paper electrophoresis and paper chromatography, included all of the amino acids commonly found in proteins. Elution of acidic, neutral, and basic amino acid and peptide fractions from C14-labelled cells was followed by measurement of total and specific radioactivities in carboxyl-C before and after acid hydrolysis. Total radioactivity was highest in proteins even 1 minute after addition of the isotope to a log-phase culture. Specific radioactivities, plotted against time, gave parallel hyperbolic curves with peptides below the curve for amino acids and with proteins still lower and almost linear. The evidence suggests rapid synthesis of peptides from amino acids.

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