A MODIFIED METHOD OF TWO-DIMENSIONAL ZONE ELECTROPHORESIS APPLIED TO MUCOPROTEINS IN SERUM AND URINE

Journal/Book: Reprinted from Nature Vol. 177 pp. 125-126 Jan. 21 1956. 1956;

Abstract: Department of Rheumatology Royal Free Hospital London DURRUM1 has described a technique of two-dimensional zone electrophoresis an paper in which the analysis in the first buffer is transferred an a strip of paper to the surface of a sheet of paper soaked in the second buffer. The strip is placed in such a position that the substances an it migrate in a direction at right angles to that in the first buffer. We wished to use two-dimensional electrophoresis to obtain direct confirmation of the positions of serum mucoprotein fractions in the serum protein pattern obtained by electrophoresis at pH 8 ··6. Two serum mucoprotein fractions may be isolated from all other serum proteins by electrophoresis at pH's between 4 · 4 and 4·· 82 (Fig. 1). Since in the latter technique filter paper is sandwiched between glass plates the following modification of Durrum's method was developed. Instead of placing the strip an the surface of the paper a slot to receive it of corresponding width and slightly greater length was cut in the centre of the second sheet at right angles to the direction of the second separation. The strip was partially dried at room temperature before being placed in position. The edges of the two pieces of paper were then carefully teased together except at either end of the strip where a gap was left. The sheet of paper had been soaked in buffer blotted and placed an the lower glass plate before receiving the strip and the flow of the new buffer into the strip concentrated the material in it as a narrow band along its centre. The absence of buffer flow inwards at either end of the strip prevented disturbance of spatial distribution along it. The upper sheet of glass was then placed over the paper the second electrophoresis carried out after a short period of equilibration and the paper-minus the strip-was then stained. ... ___MH

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