Manual

17. Optional: A reduction and alkylation of the sample is
recommended at this point in the procedure. Refer to
the ReadyPrep reduction-alkylation kit, catalog
#163-2090.
18. Store the protein extract at –70°C, apply it directly
onto an IPG strip (see Appendix for details), or proceed
to step 19.
Preparing Extracts for a MicroRotofor Run (See
Section 6 of MicroRotofor manual for alternative sample
preparation and load conditions.)
19. Prepare fresh PSB solution containing PSB diluent,
carrier ampholyte and DTT or TBP (DTT or TBP is not
required if a reduction-alkylation step is performed at
step 17). See Table 2 for recommendations.
20. One MicroRotofor run requires ~2.5 mg protein
(1 µg/µl) in a total volume of 2.5 ml. Using the above
prepared solution, prepare 2.5 ml of a 1 µg/µl dilution
of the protein extract. Load the entire 2.5 ml sample
into the MicroRotofor chamber. It may be necessary to
add extra PSB solution to fill the chamber
completely, eliminating any void volumes.
21. Run the MicroRotofor cell according to the
MicroRotofor instruction manual.
9
10005510A.qxp 8/11/2006 10:22 AM Page 9