ROUTINE HbA2 ESTIMATION BY CELLULOSE ACETATE MEMBRANE ELECTROPHORESIS゜
Two routine procedures (A and B) for estimation of HbA2 in hemolysate are described. Hemolysate (Hb concentration, about lOg/dl) is prepared by the conventional technique with carbon tetrachloride for removal of erythrocyte stroma. Both of the procedures employ cellulose acetate membrane electrophoresis with tris-EDTA borate buffer solution. A. 10 μl of hemolysate is applied to the cellulose acetate membrane (5×9 cm, pH 8.6) and an electric current (150V) is run for 90 minutes at room temperature. The electrophoresed hemoglobin stripes (A1 and A2) are eluted in Drabkin's solution, and the eluates are measured for absorbances in a spectrophotometer at 420 nm. B. Aliquot of 3 μl of hemolysate is used for the sample applied to the cellulose acetate membrane. Electrophoresis is carried out at pH 8.4 (200V, 3 mA) and 6℃ for 2 hours. The hemoglobin stripes thus appeared on the membrane are eluted in a buffer solution which is the same as that used for electrophoresis. The HbA2 content is calculated from the absorbances as shown in the equation given in the text.
