Manual

5. Add reducing agents, protease inhibitors and carrier
ampholyte as needed (Table 2)..
Table 2. Additions to PSB solution recommended
for various applications. Note that though the
applications listed often require use of chaotropes and
detergents, these agents are already included in the PSB
solution.
Protein
Component Extraction IEF Separation
MicroRotofor Cell IPG Strip
Carrier ampholyte NA 2% (w/v) 0.2% (w/v)
DTT* 50–100 mM 50–100 mM 50–100 mM
or
TBP* 2–5 mM 2–5 mM 2–5 mM
Protease inhibitor According to NA NA
manufacturer
Bromophenol Blue NA NA 0.002% (w/v)
Glycerol NA 10% NA
*Not needed if reduction-alkylation is performed at step 11.
Sample Processing
6. Weigh out 1 g of plant tissue and freeze with liquid
nitrogen.
7. Grind frozen material in liquid nitrogen with a mortar
and pestle to a fine powder.
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