Powdered acrylamide versus solution acrylamide - (Oct/01/2008 )
QUOTE (tfitzwater @ Oct 3 2008, 07:37 AM)
I used to buy 97% pure acrylamide and purify it.
Wear protective gear. Weigh 800 grams acrylamide and combine with 80 g MB3 or MB6 resin and adjust to 1 liter with Type I water.
Stir 2-3 hours (not overnight) in a 2.8 liter Fernbach flask and large magnetic stir bar. Do not heat. Do not store overnight in presence of resin. Residual catalyst from the resin will polymerize the acrylamide. Filter through a Buchner funnel using Whatman #3 over Whatman #1 into a 4 Liter sidearm flask or use a 0.45 micron disposable filter. Rinse resin. Adjust volume to 1.9 liters. Immediately rinse MB3 resin from Fernbach flask as dry resin will stick very firmly.
Add 40 g bis-acrylamide. Stir till dissolved. (2% bis is a saturated solution.) Adjust volume to 2.0 Liters with Type I water. Aliquot and store at 4°C. Stable > two years.
Clean up the area thoroughly after weighing, dissolving and filtering acrylamide.
(Vournakis, 1981. Gene Amplification and Analysis, Vol. II, p. 280.)
This method results in a > 30-fold savings over buying more highly purified liquid material. Two liters of stock is prepared in order to minimize the number of times the more risky powder must be handled. Can substitute MB1 or MB6 resin. Pharmacia (Isoelectric Focusing, 1982) says MB6 is more efficient.
Wear protective gear. Weigh 800 grams acrylamide and combine with 80 g MB3 or MB6 resin and adjust to 1 liter with Type I water.
Stir 2-3 hours (not overnight) in a 2.8 liter Fernbach flask and large magnetic stir bar. Do not heat. Do not store overnight in presence of resin. Residual catalyst from the resin will polymerize the acrylamide. Filter through a Buchner funnel using Whatman #3 over Whatman #1 into a 4 Liter sidearm flask or use a 0.45 micron disposable filter. Rinse resin. Adjust volume to 1.9 liters. Immediately rinse MB3 resin from Fernbach flask as dry resin will stick very firmly.
Add 40 g bis-acrylamide. Stir till dissolved. (2% bis is a saturated solution.) Adjust volume to 2.0 Liters with Type I water. Aliquot and store at 4°C. Stable > two years.
Clean up the area thoroughly after weighing, dissolving and filtering acrylamide.
(Vournakis, 1981. Gene Amplification and Analysis, Vol. II, p. 280.)
This method results in a > 30-fold savings over buying more highly purified liquid material. Two liters of stock is prepared in order to minimize the number of times the more risky powder must be handled. Can substitute MB1 or MB6 resin. Pharmacia (Isoelectric Focusing, 1982) says MB6 is more efficient.
Hi, May I know whether you use the same approach to prepare bisacrylamide solution?
Thx
-hkuspace graduate-
We did not purify the bis-acrylamide because it was only available at a higher purity. Limited solubility required that we add bis when we were near the final volume.
-tfitzwater-
QUOTE (tfitzwater @ Oct 23 2008, 10:38 PM)
We did not purify the bis-acrylamide because it was only available at a higher purity. Limited solubility required that we add bis when we were near the final volume.
Hi, I would like to ask whether you add bis into the stock solution of acrylamide (prepared using your way) or each time when you take some of the acrylamide stock solution out to set a gel?
THx again.
-hkuspace graduate-
Stock was 40% acrylamide/2% bis-acrylamide.
-tfitzwater-