Remazol brilliant blue as a pre-stain for the immediate visualization of human serum proteins on polyacrylamide gel disc electrophoresis

Abstract

We describe the pre-staining of proteins in normal human serum with Remazol Brilliant Blue before separation by disc electrophoresis. Critical to reproducibility are: dye concentration of 0.16 mol/L in a Tris-glycine buffer (pH 8.3), use of equal volumes of serum and dye solution, a tagging period of 2 h at room temperature, and electrophoresis of 0.1 mL of the mixture at 2.5 mA/gel for about 2 h. Advantages include speed, quality of resolution, and low background. This method was compared with Amido Black post-electrophoresis staining in 35 sera. Of these, 16 showed identical results with respect to the number of bands. In the other 19, Amido Black showed more bands in the post-transferrin region and, sometimes, in the post-albumin region. The pre-stained gels showed slower electrophoretic mobilities of the components. Protein bands eluted from pre-stained gels retained immunological reactivity.