Buffer systems for isotachophoretic separations of serum proteins

3 Buffer systems for isotachophoretic separation of serum proteins 

To get some idea what composition of buffers can be used for protein separation by isotachophoresis the following examples are presented. 

Buffer system according to Routs [176]. 0.03 M acetate, 0.011 M Tris-HCl (pH 4.5) as leading electrolyte, )3-alanine buffer (pH 9) as terminating electrolyte. 3.5% gel with 5% cross linking prepared with 0.5 mg% riboflavin, 0.03 M acetate, 0.11 M Tris (pH 4.5), 10 jul of human serum albumin (sample) is applied with an equal amount of 40% carrier ampholyte p7 5-6 on top of the gel. Isotachophoresis is carried out for 2 h at 2 mA per gel tube. 

Buffer system according to Catsimpoolas [177]. 0.24 M Tris-phosphate (pH 6.1) as leading electrolyte, 0.14 M -alanine-Tris (pH 8.0) as terminating electrolyte. 3.7% gel with 3.33% cross linking, prepared with 0.5 mg% riboflavin, 0.03 M Tris-phosphate buffer (pH 6.1), 80 jul per 1(X) ml TEMED, 3.1% sucrose. 13 cm long gels are used. Proteins are dissolved in terminating buffer and carrier ampholytes (p7 5-8, 40%) are added (35 pi to 10 pi of serum). Isotachophoresis takes 2 h at 6 mA per gel. 

Buffer system according to Davies and Rosenbaum [178]. This buffer system has been worked out for preparative purposes. 100 ml of acrylamide solution (prepared from 3.67% acrylamide, 5% bisacrylamide, in 0.012 M Tris, 0.013 M cacodylate (pH 

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