Globin chains analysis: improved resolution by electrophoresis in urea-acetic acid-Triton X-100

Abstract

To improve the resolution and rapidity of globin chains separation, we have modified the basic technique of globin chain electrophoresis in urea-acetic acid-Triton X-100. Haemolysates from anticoagulated cord or adult blood samples were submitted to urea-acetic acid-Triton X-100 polyacrylamide gel electrophoresis using a 15% polyacrylamide gel cast in a mini slab cell which allows a rapid analysis of globin chains samples. After staining proteins with Coomassie brilliant blue R-250, the relative amounts of globin chains were determined by scanning. This new procedure has allowed us to obtain a better separation of the normal and abnormal globin chains than described previously. All the normal globin chains, i.e. A gamma, G gamma, delta, beta and alpha, are well separated by this modified technique. Semi-quantification of the G gamma/A gamma ratio has been performed. This simple and rapid method is also suitable for the global identification of the globin chain involved in the most common abnormal haemoglobin variants, except beta-S.