TRIS Buffer for the Demonstration of Haemoglobin A₂by Paper Electrophoresis

Abstract

Trishydroxymethylaminomethane (TRIS) buffer has been recommended for finer separation of serum proteins in paper electrophoresis (Aronsson and Gronwall, 1957). We have employed it for the examination of haemoglobin variants and have found that, with a slight modification, it is suitable for this purpose. When used with a vertical tank the fractions do not move as far from the point of application as they do with barbiturate buffer (pH 8.6), but they are more sharply defined and the bands are narrower. In view of this it is possible to recognize, with the naked eye, small fractions which would escape discovery on paper electrophoresis with barbiturate buffer. This is particularly useful for the examination of specimens for haemoglobin A2. Even normal proportions of haemoglobin A2 are easily visible, and abnormal proportions of haemoglobin A2 are recognized at once. By this technique the non-haemoglobin protein X of Derrien (1959), which is present in some haemolysates, separates clearly from haemoglobin A2, and on FIG. la