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Human haptoglobin phenotypes on native-page using tetramethylphenylenediamine (tmpd) staining

Authors:Marghoob Hasan, Abdelmarouf Mohieldein, Mohammad Alzohairy, Moinuddin khan
Int J Biol Med Res. 2012; 3(1): 1342-1344  |  PDF File


This paper describes staining method for the H2O2-peroxidase activity of Hb-Hp complex (Hemoglobin-Haptoglobin) on polyacrylamide electrophoresis gel using N,N,N′,N′-tetramethyl-p-phenylenediamine. Haptoglobin phenotyping is performed for many decades in biomedical research due to its relevant association with many clinical conditions. Various advanced techniques are available; however, peroxidase activity based staining is still in use. We evaluated the use of an alternative stain for peroxidase staining of Hb-Hp complex. This can be used safely in routine procedure for the identification of haptoglobin polymorphism. N,N,N′,N′-tetramethyl-p-phenylenediamine (TMPD) acts as an effective indicator of peroxidase activity on polyacrylamide electrophoresis gels. The test is easy to perform, rapid, sensitive, and reliable. The procedure produces vivid bright blue bands (Wursters blue) on a clear background.