IPG rehydration buffer
To prepare 50 ml of rehydration buffer, dissolve 25.0 g of urea in deionized H2O and adjust the volume to 50 ml. Add 0.5 g of Serdolit MB-1 mixed-bed ion-exchange resin (Serva), stir for 10 minutes, and filter. Add 0.5 g of CHAPS, 0.2 g of DTT, and 1.25 ml of Pharmalyte (pH range 3-10) to 48 ml of the urea solution.
Note:
For IEF of hydrophobic proteins, 6 M urea with 2 M thiourea is superior to 8 M urea. For IEF of basic proteins, the use of DeStreak instead of IPG rehydration buffer is preferable.
