SDS PAGE: problem with acrylamine and bisacrylamide - (Mar/17/2008 )
I have just started working with SDS PAGE. but when i dissolve bisacrylamide (1%) after dissolving acrylamide (29%) the solution becomes turbid and gel does not solidify. All the other chemicals TMED, APS , and other buffere solutions are working properly. Bisacryamide and acrylamide both are not expired. Can anyone tell why it is so and how to overcome this problem?
Just stir it for long time.
set the solution in a pan of warm water while stirring, it will dissolve faster.
When preparing acryl-bis solution let it stir for more than 30 min, always use an ambar bottle for storage and use it within a month. If a gel don't polimeryze could be: not enough TEMED and ammonium persulfate, reagents too old. The APS should be prepare as fresh as posible. I usually prepare small quantity and use within a week (store at 4C). Let the gel polymerize at least 1 hour. If preparing a discontinue gel when adding the buthanol use just a thin layer and wash it with ddH2O (alcohol will inhibit the polymerization of the stacking gel).