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4. Vortex periodically and incubate at room temperature

until you have a clear solution (2–3 min).

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

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 13.

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