Distribution of Iron-59 in Stroma-free Thalassæmic Erythrocytes examined by means of Gel-filtration

Abstract

A NUMBER of methods are available for examining the distribution of iron incorporated into the soluble and other portions of the immature erythrocyte. Tishkoff has utilized precipitation of the hæmolysate by trichloroacetic acid (TCA)(ref. 1). Allen and Jandl², and Greenough et al.³, used ion exchange chromatography on ‘IRC 50’. Lothe and Falbe-Hansen separated their non-hæm iron from hæmoglobin electrophoretically⁴. Other laboratories isolated hæmin from the hæmolysate by Fisher's method⁵. However, all the aforementioned procedures involve one or more drastic steps detrimental to the physical state of various components in the erythrocyte (destruction of secondary and/or tertiary structure of proteins, changes in the dissociation of ionizable molecules and so on). To overcome this difficulty, we used gel-filtration on polymerized dextrans, introduced first by Porath and Flodin⁶, with satisfactory results.