Indices of membrane alterations in beta-thalassemic erythrocytes

Analytical protocols for the study of thalassemic erythrocyte membrane alte rations are described, Denatured hemoglobin derivatives and aggregated band 3 are separated from detergent membrane extracts by gel-filtration as high -molecular-weight aggregates and quantitated spectrophotometrically. Membra ne-bound, low-molecular-weight iron is measured on SDS-solubilized ghosts b y a ferrozine-based colorimetric test. We adapted these methods for microsc ale preparation and analysis of erythrocyte ghosts in order to have suitabl e tools to estimate oxidative membrane damage in human samples, Data from 1 1 beta- thalassemia intermedia patients and from 10 normal controls are rep orted as an example of the application of these methods.