Hoefer IEF100 - Manual (Page 59)

• p53
Recipes
IEF Sample Extraction Buffer for 2D
1A. Urea Sample Buffer Solution
Prepares 25 ml
9.5 M urea, 4% CHAPS
Final Concentration Amount
Urea (FW 60.06) 9.5 M 14.26 g
CHAPS 4% (w/v) 1.0 g
Deionized water to 25 ml
Store in 1 ml aliquots at -20 °C or below.
Prior to use, add 6 mg/ml DTT to get a final sample buffer composition of
40 mM DTT.
Optional: Add carrier ampholytes, such as SERVALYTS, to a concentration of
2% v/v (20 μl per ml sample buffer solution).
— OR —
1B. Thiourea + Urea Sample Preparation Solution
Prepares 25 ml
7 M urea, 2 M thiourea, 4% CHAPS, 40 mM DTT
Final Concentration Amount
Urea (FW 60.06) 7 M 10.51 g
Thiourea (FW 76.12) 2 M 3.8 g
CHAPS 4% (w/v) 1.0 g
Deionized water to 25 ml
Store in 1 ml aliquots at -20 °C or below.
Prior to use, add 6 mg/ml DTT to get a final sample buffer composition of
40 mM DTT.
Optional: Add carrier ampholytes, such as SERVALYTS, to a concentration of
2% v/v (20 μl per ml sample buffer solution).