Hoefer SE900 - Manual (Page 31)

• p25
Solutions
1. Acrylamide Stock Solution
(30.8% T 2.6% C Bis, 1000 ml)
Acrylamide
(FW 71.08) 30% w/v 300 g
Bis*N,N' Methylenebisacrylamide (FW 154.2) 0.8% w/v 8 g
Deionized H
2
O to 1000 ml.
Store at 4 °C away from light.
2. 1.5 M TrisCl, pH 8.8
(4X Resolving gel buffer, 2 liter)
Tris
(FW 121.1) 1.5 M 363.3 g
Disolve into ~1.5 liters deionized H
2
O.
4 N HCl (4N hydrochloric acid) to pH 8.8.
Deionized H
2
O to 2000 ml.
Store at 4 °C.
3. 10% w/v SDS Solution
(100 ml)
Sodium dodecylsulfate (SDS)
(FW 288.4) 0.35 M 10 g
Deionized H
2
O to 100 ml.
Store at room temperature.
4. 10% w/v APS
(Initiator, 1 ml)
Ammonium persulfate (APS)
(FW 228.2) 0.44 mM 5 g
Deionized H
2
O to 5 ml.
Prepare just prior to use.
Fresh APS “crackles” when water is added. If yours does not, replace it with
fresh stock.
5. 10% v/v TEMED
(Catalyst, 5 ml)
N, N, N’, N’-Tetramethylethylene-
diamine (TEMED)
(FW116.2 ) ~0.65 M 0.5 ml
Deionized H
2
O to 5 ml.
Prepare just prior to use.
Store in a dark glass bottle at room temperature away from light.
TEMED is flammable and should be dispensed in a fume hood.
Caution! Acrylamide is a neurotoxin. Extreme
care should be used when handling and
preparing acrylamide solutions.