Properties of thin-rod immobilized pH gradients.
Immobilized pH gradient gel rods, 1.5 mm in diameter, were cast with a manifold connected to high-precision burettes. The reproducibility of gel length was ca. 1.7 mm. The average standard deviation sigma x for spot position was 2 mm after one-dimensional and 5.8 mm after two-dimensional runs. In order to bring to completion the elution of the salt fronts into the electrode compartments, carrier ampholytes had to be included in the gel formulation at concentrations of at least 0.5-1%, depending on the pH range. The presence of carrier ampholytes, however, was troublesome in two respects: the gel tended to shrink and the cathodic bands drifted with time. Ionic components in the sample were tolerated up to the following concentrations: NaCl 8 mumoles, sodium dodecyl sulfate 10 micrograms per tube. In presence of non-ionic detergents, the gels moved as a whole towards the cathode.
Gianazza
E
E
Chair of Biochemistry, Faculty of Pharmacy, University of Milano, Italy.
Tedesco
G
G
Cattò
N
N
Bontempi
L
L
Righetti
P G
PG
D000665:Ampholyte Mixtures; D002021:Buffers; D003902:Detergents; D004586:Electrophoresis; D004864:Equipment and Supplies; D005782:Gels; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D011506:Proteins / Q000032:analysis; D015203:Reproducibility of Results; D012492:Salts; D012967:Sodium Dodecyl Sulfate; D014867:Water
D000665:Ampholyte Mixtures; D002021:Buffers; D003902:Detergents; D005782:Gels; D011506:Proteins; D012492:Salts; D014867:Water; D012967:Sodium Dodecyl Sulfate