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. Aγ, Gγ, δ, β and α, are well separated by this modified technique. Semi-quantification of the Gγ/Aγ 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 β-S.