Electrophoresis in non-dissociating buffer systems

For the discontinuous buffer systems, the three widely used protocols, i.e., for high pH (Davis, 1964 Ornstein, 1964), neutral pH (Williams and Reisfeld, 1964) and low pH (Reisfeld et al., 1962), are in Table 16.5. The neutral buffer system has a poor buffering capacity and is only useful for proteins which are unstable in the high pH system. The low pH system is used for basic proteins (histones, several of the POase isozymes). It is imperative to check carefully the pH of all solutions. 

Solutions Stacking gel Volumes (ml) required for resolving gel with polyacrylamide concentration (%) of  

The stacking gel and resolving gel buffers are the same as in the Laemmli system (Table 16.3). 

Stacking gel buffer dissolve 4.9S g Tris in 40 ml distilled water and titrate to pH 

5 with I M orthophosphoric acid. Complete to 100 ml with distilled water. Resolving gel buffer dissolve 6.85 g Tris in 40 ml distilled water and titrate to pH 

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