Manual

Note: Following fractionation with the MicroRotofor, it is
recommended to perform an SDS-PAGE analysis profiling
all 10 fractions. This will illustrate the protein content of
each fraction. See the Appendix for recommendations
pertaining to SDS-PAGE analysis of MicroRotofor fractions.
For subsequent analysis of MicroRotofor fractions by 2-D
PAGE, the ampholyte concentration in samples should not
exceed 0.2–0.5%. If fractions contain high amounts of
protein, dilution prior to loading onto the IPG strip (by 1:10
or greater) will be sufficient to reduce the ampholyte
concentration. In cases where protein levels are lower, use
of the ReadyPrep 2-D cleanup kit (catalog #163-2130) for
ampholyte removal is recommended.
Section 5
Appendix
Preparation for SDS-PAGE
CHAPS, a component of the PSB diluent, may interfere
with SDS-PAGE. Remove CHAPS from the extracts (for
example, with the ReadyPrep 2-D cleanup kit) or dilute the
extracts 1:1 with 1x Laemmli buffer prior to SDS-PAGE.
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