Gradient Master - Wolf Laboratories

Figure 3. Loading volume influences band width; 5-20% gradient. While the 0.1 and 0.25 ml
samples gave identical peak shapes, the 0.5 and 0.75 ml sample volumes clearly lead to a broadening
of the band and the resultant loss of resolution. Thus, for the SW41 rotor tubes, sample volumes in
5-20% gradients must be limited to 0.25 ml and fairly short runs if high resolution is required. These
gradients were fractionated using the hand crank (pre-motorized).
A final limitation of these gradients will become evident when examining the 5-45% profiles
below. The range of S values that can be separated on a given gradient is limited here because the
low viscosity and density of the 20% end point does not retard the sedimentation of large particles.
In Figure 1, for example, particles with an S value of 250-750S would have been separated adequately
but 1000S particles would have pelleted in this run.
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