Hoefer IEF100 - Manual (Page 60)

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IPG Strip Rehydration Solution
2A. Urea Rehydration Stock Solution
Prepares 50 ml
8 M urea, 2% CHAPS, 0.002% bromophenol blue
Final Concentration Amount
Urea (FW 60.06) 8 M 24 g
CHAPS 2% (w/v) 1.0 g
Bromophenol blue 0.002% 1 mg
Deionized water — to 50 ml
Store in 3 ml aliquots at -20 °C or below. 3 ml is sufficient to rehydrate six
24 cm IPG strips.
Just prior to use for IPG strip rehydration:
• Add 0.5 – 2.0% (v/v) carrier ampholytes (SERVALYTS).
• Add 9 mg DTT per 3 ml aliquot of rehydration stock solution 20 mM DTT.
• Protein sample can also be added to the 3 ml of rehydration solution.
2B. Thiourea Rehydration Stock Solution
Prepares 50 ml
7 M urea, 2 M thiourea, 2% CHAPS, 0.002% bromophenol blue
Final Concentration Amount
Urea (FW 60.06) 7 M 21 g
Thiourea (FW 76.12) 2 M 7.6 g
CHAPS 2% (w/v) 1.0 g
Bromophenol blue 0.002% 1 mg
Deionized water — to 50 ml
Store in 3 ml aliquots at -20 °C or below. 3 ml is sufficient to rehydrate six
24 cm IPG strips.
Just prior to use for IPG strip rehydration:
• Add 0.5 – 2.0% (v/v) carrier ampholytes (SERVALYTS).
• Add 9 mg DTT per 3 ml aliquot of rehydration stock solution 20 mM DTT.
• Protein sample can also be added to the 3 ml of rehydration solution.