Bio-Rad MicroRotofor™ Cell Lysis Kits User Manual
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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.
Preparation for IEF on an IPG Strip
Following step 17, the sample extract can be loaded onto
an IPG strip after appropriate dilution. See Table 3 for
recommendations on how much protein sample to load
onto an IPG strip. Dilution of the sample can be done using
protein solubilization buffer (PSB) as a rehydration/sample
buffer. However, some critical components need to be added
to the PSB solution to make it IEF-compatible (Table 2).
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