Which electrodic solutions for immobilized pH gradients?
Carrier ampholytes covering a pH range corresponding to, or narrower than, the span of the immobilized pH gradient (IPG) are a most suitable electrodic solution for IPGs. They are able to collect, and completely remove from the gel, much higher amounts of non-buffering ions than are solutions of acidic and basic amino acids. This makes it possible to directly run IPGs just after their polymerization, without the need of a washing step to remove catalysts and unreacted Immobiline monomers. The same applies most advantageously when the gel formulation includes urea and/or detergents. Ions contributed by the sample solution are also prevented from casting high-conductivity ridges around the electrodes, without any need either of a dialysis step or of an increased slab size with pH plateaus. The migration of the sample proteins toward their equilibrium position is faster in the presence of carrier ampholytes. The effective concentrations of the latter are in the range 0.3-1%.
Gianazza
E
E
Quaglia
L
L
Caccia
P
P
Righetti
P G
PG
D000665:Ampholyte Mixtures; D003902:Detergents; D004553:Electric Conductivity; D004566:Electrodes; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing / Q000379:methods*; D012996:Solutions; D014508:Urea
D000665:Ampholyte Mixtures; D003902:Detergents; D012996:Solutions; D014508:Urea