We developed a novel polyacrylamide gel electrophoresis (PAGE) method to stack and separate human hemoglobins (Hbs) based on the concept of moving reaction boundary (MRB),differing from he classic isotachophoresis (ITP)-based stacking PAGE in the aspect of buffer composition,including the electrode buffer (pH 8.62 Tris-Gly),sample buffer (pH 6.78 Tris-Gly) and separation buffer (pH 8.52 Tris-Gly).